Toxoplasma gondii prevalence and assays used in different groups of domestic animals in the Czech Republic until year 2000
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More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
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Toxoplasmosis is a significant zoonosis that affects humans and warm blooded animals. The definitive hosts of parasite Toxoplasma gondii are cats and other felids. Many species of domestic, wild or zoo animals may serve as intermediate hosts.
In humans, clinical form of toxoplasmosis is rare in immunocompetent people, while it may leads to eye diseases, CNS or generalized infection in immunocompromissed individuals as well as interfere with the course or outcome of pregnancy. In Europe, T. gondii seroprevalence in humans ranges from 8% to 77% (Dubey 2010). In the Czech Republic, T. gondii antibodies were detected in 35% and 25% pregnant women by Sabin-Feldman Test (SFT) and Complement Fixation Test (CFT), respectively (Hejlicek et al. 1999). Repeated prevalence studies in humans in some European countries (France, Belgium, Sweden and Norway), revealed an evident trend of a decrease in T. gondii seroprevalence (Welton and Ades 2005). The same trend is observed in the Czech Republic. The prevalence of infection varies among ethnic groups due to sanitary and cooking habits. Consumption of raw or almost raw, dried, cured or smoked meat from domestic animals, unpasteurized goat milk or consumption of meat from wild animals may be associated with ingestion of the parasite (Kijlstra and Jongert 2008, Jones et al. 2009). Higher prevalence was found also in people who had frequent contact with animals and soil, such as abattoir workers, garbage handlers and waste pickers (Dubey and Beattie 1988). Children playing with dogs and cats can be infected by direct contact because animals can act as mechanical vectors (Etheredge et al. 2004).
In animals, T. gondii infection is a frequent cause of early embryonic death and resorption, fetal death and mummification, abortion, still birth and neonatal death. Thus, toxoplasmosis in domestic and farm animals is a disease of great importance for veterinary medicine and husbandry since it can cause productive and economic losses.
T. gondii antibodies have been found in animals worldwide. Seroprevalence to T. gondii varies among countries, within different areas of a country and within the same city. Dubey (2010) summarized the results of seroprevalence studies performed on different groups of animals from several countries.
In the Czech Republic, some important studies concerning T. gondii in animals were done in past years. The seroprevalence of T. gondii infection in domestic animals obtained by different serological methods is summarized in Table 1.
Animal | Prevalence | Assay | Reference |
Cat | 17 – 91% | SFT, CFT, MPA, IFAT | Havlik and Hubner 1958, Zastera et al. 1966, Zastera et al. 1969, Svoboda and Svobodova 1987, Svoboda 1988 |
Dog | 15 – 58% | SFT, CFT, MPA | Havlik and Hubner 1958, Zastera et al. 1966, Zastera et al. 1969, Svoboda and Svobodova 1987, Hejlicek et al. 1995, Hejlicek et al. 1995 |
Sheep | 4 – 77% | SFT, CFT, MPA, IHA, IFAT | Zastera et al. 1966, Zastera et al. 1969, Arnaudov et al. 1976, Kozojed et al. 1977, Hejlicek and Literak 1994c |
Goat | 20 – 86% | SFT, CFT, IFAT | Havlik and Hubner 1958, Zastera et al. 1966, Zastera et al. 1969, Hejlicek and Literak 1994b, Literak et al. 1995, Slosarkova et al. 1999 |
Cattle | 2 – 42% | SFT | Havlík and Hubner 1958, Zastera et al. 1969, Kozojed et al. 1977 |
Pig | 0.1 – 38% | SFT, CFT, MPA | Havlik and Hubner 1958, Zastera et al. 1969, Kozojed et al. 1977, Hejlicek and Literak 1993, Hejlicek and Literak 1994b, Vostalova et al. 2000 |
Horse | 4 – 11% | SFT, CFT | Havlik and Hubner 1958, Zastera et al. 1969, Kozojed et al. 1977, Hejlicek and Literak 1994a, Zastera et al. 1966 |
Gallinaceous bird | 0 – 20% | SFT | Zastera et al. 1965, Zastera et al. 1969, Kozojed et al. 1977, Literak and Hejlicek 1993 |
Water fowl | 2 – 33% | SFT | Zastera et al. 1965, Zastera et al. 1969, Literak and Hejlicek 1993 |
Rabbit | 6 – 95% | SFT, CFT | Havlik and Hubner 1958, Havlik and Hubner 1960, Zastera et al. 1969 , Kunstyr et al. 1970, Hejlicek and Literak 1994d |
Toxoplasma gondii prevalence and assays used in different groups of domestic animals in the Czech Republic until year 2000
In a group of game animals, a prevalence of 15% was found in wild boars by SFT (Hejlicek et al. 1997), 4 – 31% in hares by SFT or Microprecipitation in Agar (MPA) (Havlik and Hubner 1958, Zastera et al. 1966, Vosta et al. 1981, Hejlicek et al. 1997), and 14 – 58% prevalence in wild ruminants by SFT (Havlik and Hubner 1958, Zastera et al. 1966, Hejlicek et al. 1997).
These studies were performed by one or by a combination of methods such as SFT, CFT, MPA and Indirect Hemaglutination Assay (IHA). Nowadays, these methods are less frequently used; it is preferred to use Modified Agglutination Test (MAT), and/or Indirect Fluorescent Antibody Test (IFAT), and/or an Enzyme-Linked Immunosorbent Assay (ELISA), and/or a Latex Agglutination Test (LAT). This trend is also evident from a recent review summarized worldwide prevalence of T. gondii infection in animals and humans (Dubey 2010).
Based on the results of examination of different groups of animals in the State Veterinary Institute Prague in years 2003 – 2006, it is evident that lethal toxoplasmosis in the Czech Republic is the most important in some species of zoo animals; while in domestic animals it was not proved (Sedlak and Bartova 2007). Contrary, the sera of cats and dogs were the most frequently examined. Insufficient attention is paid to small ruminants that can abort or have reproduction disorders due to toxoplasmosis with subsequent economic losses.
That is why during the last 10 years, our research team focused on T. gondii serosurveys in different groups of animals to obtain actual data and to evaluate which group of animals is the most affected by T. gondii infection. Following parts of chapter summarises the results of seroprevalence studies in domestic, game and zoo animals tested by using IFAT, ELISA and LAT with the possibility to compare the results with those obtained from other countries with the same methods used. The results of experimental studies and cases of clinical toxoplasmosis recorded in the Czech Republic are mentioned too.
Serological studies
During years 1995-2012, the samples of blood were collected from different groups of animals and examined for specific T. gondii antibodies. The animals tested for T. gondii antibodies were clinically healthy, no case of abortion or other symptoms of toxoplasmosis were recorded. The blood samples were collected by veterinarians on farms, zoo or during hunting seasons and sent to State Veterinary Institute Prague for routine examination.
In a group of domestic animals, in total 4254 animals were tested with the following number of animals used: 286 cats, 413 dogs, 547 sheep, 251 goats, 546 cattle, 551 pigs, 552 horses and 1108 poultry (217 chickens and 293 broilers, 60 turkeys, 178 geese and 360 ducks). The animals came from 2 – 14 different districts of the Czech Republic (Figure 1).
Map of the Czech Republic showing the sampled area with domestic and game animals tested for T. gondii antibodies.
Sera of domestic animals were tested for T. gondii antibodies by an indirect fluorescent antibody test (IFAT), using the Sevatest Toxoplasma Antigen IFR (Sevac, Prague, Czech Republic) and specific conjugates, by an ELISA (Institut Pourquier, Montpellier, France), or by a latex agglutination test (Pastorex TM Toxo, Biorad, France). The data on the method and cut‐off used, specific conjugate for IFAT and producer are summarized in Table 2.
In a group of domestic animals, T. gondii antibodies were found in 66% goats, 59% sheep, 44% cats, 36% pigs, 26% dogs, 23% horses, 12% poultry (43% goose, 14% ducks, and 0.3% in broiler; turkeys and chickens were negative) and 9.7% cattle. The results of serological examination including the number of samples tested, the method and cut-off used, the number and percentage of positive samples, titres or %S/P obtained in positive samples and reference about published data are summarized in Table 3.
Animal | Assay (cut-off) | Conjugate for IFAT | Producer |
Cat | IFAT (≥40) | anti-cat IgG | Sigma Aldrich, USA |
Dog | IFAT (≥40) | anti-dog IgG | Sigma Aldrich, USA |
Sheep | ELISA (≥50%S/P) | – | Sigma Aldrich, USA |
Goat | ELISA (≥50% S/P) | – | Sigma Aldrich, USA |
Cattle | ELISA (≥50% S/P) | – | Sigma Aldrich, USA |
Pig | ELISA (≥50% S/P) | – | Sigma Aldrich, USA |
Horse | LAT | anti-horse IgG | VMRD, Pulman, USA |
Chicken Broiler | IFAT (≥40) | anti-chicken IgG | Sigma Aldrich, USA |
Turkey | IFAT (≥40) | anti-chicken IgG | Sigma Aldrich, USA |
Goose | IFAT (≥40) | anti-duck IgG | KPL, USA |
Duck | IFAT (≥40) | anti-duck IgG | KPL, USA |
Serologic method, cut-off, specific conjugates for IFAT and producer used in domestic animals.
Animals | T. gondii | Assay (cut-off) | Titres or %S/P | Reference | ||
n | positive | % | ||||
Cat | 286 | 126 | 44 | IFAT (40) | 40 – 81920 | Sedlak and Bartova 2006b |
Dog | 413 | 107 | 26 | IFAT (40) | 40 – 10240 | Sedlak and Bartova 2006b |
Sheep | 547 | 325 | 59 | ELISA (50%S/P) | 50 – 200 | Bartova et al. 2009a |
Goat | 251 | 166 | 66 | ELISA (50%S/P) | 56 – 191 | Bartova et al. 2012 |
Cattle | 546 | 53 | 9.7 | ELISA (50%S/P) | 50 – 200 | Bartova et al. (unpublished) |
Pig | 551 | 198 | 36 | ELISA (50%S/P) | 50 – 337 | Bartova and Sedlak 2011 |
Horse | 552 | 125 | 23 | LAT | – | Bartova et al. 2010a |
Poultry | ||||||
Chicken | 510 | 0 | 0 | IFAT (40) | – | Bartova et al. 2009a |
Broiler | 293 | 1 | 0.3 | IFAT (40) | 40 | Bartova et al. 2009a |
Turkey | 60 | 0 | 0 | IFAT (40) | – | Bartova et al. 2009a |
Goose | 178 | 77 | 43 | IFAT (40) | 40 – 2560 | Bartova et al. 2009a |
Duck | 360 | 52 | 14 | IFAT (40) | 40 – 320 | Bartova et al. 2009a |
The result of serological examination of domestic animals with method used, cut-off, titres and %S/P in positive samples
Experimental studies
In the Czech Republic, two experimental studies were conducted on domestic poultry.
The first study was conducted on chickens (Gallus domesticus) that were inoculated per os with two different doses of T. gondii oocysts (Sedlak et al. 2000b). Antibodies to T. gondii were detected by IFAT first on day 14 p.i.; all chickens were serologically positive on days 21 and 28 p.i. No clinical symptoms were recorded. Parasite T. gondii was isolated from heart, muscle, spleen and brain. In one case, no T. gondii was isolated from any organ. Based on this experiment chickens seems highly resistant to T. gondii infection.
The second experimental study was conducted on domestic ducks (Bartova et al. 2004). Ducks were inoculated per os with different doses of T. gondii oocysts. Antibodies to T. gondii were detected in all ducks by IFAT on day 7 p.i. Antibody titres were found in the range of 20–640 depending on the infectious dose of the oocysts. From day 14 p.i., antibody titres increased to 80–20 480. Bioassay in mice revealed T. gondii in the breast and leg muscles, heart, brain, liver and stomach. The infected ducks showed no clinical symptoms, however, the results of bioassay indicate that, compared to gallinaceous birds, domestic ducks are relatively susceptible to T. gondii infection.
Cats
Clinical signs of toxoplasmosis in cats include fever, anorexia, dyspnea, uveitis, pneumonitis and others. Kittens can develop acute toxoplasmosis and die from it. The seropositivity increases with the age of cat, indicating postnatal transmission of infection. T. gondii antibodies have been found worldwide (Dubey 2010). Seroprevalence varies among countries, within different areas of a country and within the same city. In Europe, the highest prevalence 76% was found by SFT in Turkey (Karatepe et al. 2008), while the lowest 17% in Israel by ELISA (Salant and Spira 2004). In the Czech Republic, we found 44% prevalence by IFAT. In the previous studies from the Czech Republic, 17% – 91% prevalence was found by SFT, CFT and MPA. During the last 20 years, there is a trend of decreasing seroprevalence especially in cats staying at home and fed with commercial diet.
Dogs
T. gondii antibodies have been found in canine sera worldwide. Seroprevalence increases with age indicating postnatal infection, is higher in dogs from rural areas, in dogs housed exclusively outdoors, in dogs eating birds, small mammals, meat, viscera and home-cooked meals (Lopes et al. 2011b). In Europe, the highest prevalence 75% was found by SFT in Turkey (Aktas et al. 1998), while the lowest 5% in Sweden by ELISA (Lunden et al. 2002). In the Czech Republic, we found 26% prevalence by IFAT. In the previous studies from the Czech Republic, 4% – 58% prevalence was found by SFT or CFT. The lower prevalence is recorded in dogs staying at home and fed with commercial diet.
Sheep
T. gondii has been recognized as one of the main cause of infective ovine abortion in New Zealand, Australia, the United Kingdom, Norway and the United States. In the Czech Republic, not yet a case of toxoplasmic abortion has been recorded in sheep herds. Seroprevalence was shown to increase with age, suggesting that animals acquire infection postnatally, however transplacental transmission of T. gondii may be more common than previously believed. Antibodies to T. gondii have been found in sheep worldwide (Dubey 2010). There is no validation of any serological test for the detection of T. gondii infection in sheep; different methods and cut-off are used.
In Europe, the highest prevalence 96% was found by ELISA in Turkey (Mor and Arslan 2007), while the lowest 10% was found in Slovak Republic by SFT (Kovacova 1993). We found 59% prevalence by ELISA. In the Czech Republic, 4% – 77% prevalence was found in past years.
Based on experimental studies, T. gondii was more frequently detected in brain and heart than in muscles; however T. gondii was detected also in milk (Camossi et al. 2011). Attention should be paid to meat or milk consumed without sufficient temperature treatment.
Goats
T. gondii antibodies have been found in goats worldwide (Dubey 2010). In Europe, the highest prevalence 91% was found by LAT in Netherland (McSporran et al. 1985), while no antibodies were found in Poland by IFAT (Gerecki et al. 2005). We found 66% prevalence by ELISA. In the Czech Republic, 20% – 86% prevalence was found in past years.
Goats appear to be more susceptible to clinical toxoplasmosis compared to other domestic animals, and even adult goats could die of acute toxoplasmosis. In the Czech Republic, toxoplasmosis was diagnosed in two Angora goat herds in South Moravia with an outbreak of abortions and births of weak kids; the goats showed also iodine deficiency (Slosarkova et al. 1999). Based on several experimental studies conducted on goats, T. gondii was detected in liver, muscles, heart, diaphragm, brain, kidneys and could be excreted in semen and milk. Attention should be paid to raw goat meat and milk if consumed without sufficient temperature treatment.
Cattle
Serum antibodies to T. gondii have been found in cattle in many surveys worldwide (Dubey 2010). In Europe, the highest seroprevalence 92% was found by MAT in Italy (Avezza et al. 1993), while no antibodies were found in Slovak Republic (Pleva et al. 1997) and Turkey (Oz et al. 1995). Actual prevalence rates are likely to be lower than indicated because of problem with the specifity of the tests used. The SFT test gives false or erratic results with cattle sera; on the other hand a titer of 1:100 or higher in the MAT appears to be indicative of T. gondii infection in cattle (Dubey 2010). We found 9.7% seroprevalence by ELISA. In the previous studies in the Czech Republic, 2% – 42% seroprevalence was found by SFT and DT.
There are no confirmed reports of clinical toxoplasmosis in adult cattle. In cattle, T. gondii can be transplacentally transmitted resulting in aborts; but it is probably a rare occurrence. There is more important parasite Neospora caninum leading to abortion in cattle. In the Czech Republic, there is very low prevalence of N. caninum in herds of cattle. The ingestion of beef or dairy products is not considered important in the epidemiology of T. gondii because cattle are not a good host for this parasite. Attempts to isolate T. gondii from cattle tissues have been unsuccessful, that is why it does not present risk of infection for humans.
Pigs
Clinical manifestation of toxoplasmosis in pigs could include diarhea, encephalitis, pneumonitis, necrotic hepatitis and abortion. Surveys based on the presence of T. gondii antibodies in blood sera of pigs have been reported worldwide (Dubey 2010). In Europe, T. gondii prevalence declined in the last decade especially because of good management system. There is a different sensitivity and specifity of the assays used for serosurveys in the following order MAT, IHA, LAT and ELISA starting with the most sensitive one. Good correlation was obtained between ELISA and MAT. In Europe, the highest prevalence 64% was found by IFAT in Italy (Genchi et al. 1991), while only 1% prevalence was found by the same method used in Austria (Edelhofer 1994). In the Czech Republic, we found 36% prevalence by ELISA. In the previous studies from the Czech Republic, 0 – 38% prevalence was found by SFT, CFT or MPA.
The higher prevalence is found among pigs from small backyard operations, while the prevalence among pigs from traditional large farms and modern large-scale farms is usually lower. Attention should be paid if pork meat is consumed nearly raw or without sufficient temperature treatment.
Horses
Horses have been shown to be susceptible to Toxoplasma infection (Tassi 2006) however there is no confirmed report of clinical toxoplasmosis. Serum antibodies to T. gondii have been found in horses in many surveys worldwide (Dubey 2010). In Europe, the highest prevalence 37% was found by SFT in Turkey (Gazayagci et al. 2011), while the lowest 1% in Sweden by DAT (Jakubek et al. 2006). In the Czech Republic, we found 23% by LAT. In the previous studies from the Czech Republic, 4 – 11% prevalence was found by SFT or CFT.
By reason that equine meat represents an important source of food in many human communities, infected equine meat could represent potential risk of T. gondii infection for humans.
Poultry
In general, there is a different sensitivity of birds to T. gondii infection. Owls and other predatory birds and domestic poultry seem to be resistant to T. gondii infection, while e.g. rock partridge (Alectoris graeca), pigeons and canaries are highly susceptible to toxoplasmosis. In Europe, there were some reports of birds (galliformes, columbiformes, psittaciformes and passeriformes) that died due to toxoplasmosis (Dubey 2010). Toxoplasmosis can also lead to drop in egg production and high mortality in embryonated eggs. In the Czech Republic, confirmed clinical toxoplasmosis has not been recorded in birds. Little is known concerning the validity of the serologic tests for the detection of T. gondii antibodies in avian sera. It is preferred to use MAT, nevertheless other methods such as SFT, CFT, ELISA and IFAT have been used worldwide.
We found higher prevalence in water fowls (43% and 14% in goose and ducks, respectively) compared to gallinaceous poultry (0.3% in broiler; turkeys and chickens were negative). In Europe, higher prevalence 36% was found in chicken from Austria by MAT (Dubey et al. 2005), or 20% in turkeys by ELISA in Germany (Koethe et al. 2011).
T. gondii have been isolated from brain, heart and leg muscles, but not from the pectoral muscle and liver (Dubey et al. 1993).
Serological. studies
In majority of game animals, the course of infection is subclinical. However, considering the high prevalence of T. gondii infection in game animals, they should be taken into account as the possible source of infection for human.
A total of 1618 game animals were tested, including 720 wild ruminants or ruminants living in reservations (377 red deer, 79 roe deer, 14 sika, 143 fallow deer, 105 mouflon and 2 reindeer), 565 wild boars and 333 hares. The animals came from 3 – 11 districts of the Czech Republic (Figure 1).
Sera of game animals were tested for T. gondii antibodies by an IFAT, using the Sevatest Toxoplasma Antigen IFR (Sevac) and specific conjugates (Table 4). Sera with titer ≥40 were marked as positive.
Animal | Conjugate for IFAT | Producer |
Wild boar | anti-swine Ig G | Sigma, Praha |
Hare | anti-rabbit Ig G | Sigma Aldrich, USA |
Red deer | anti-deer Ig G | KPL Inc. Maryland |
Sika | anti-deer Ig G | KPL Inc. Maryland |
Fallow deer | anti-deer Ig G | KPL Inc. Maryland |
Roe deer | anti-deer Ig G | KPL Inc. Maryland |
Mouflon | anti-goat Ig G | VMRD, USA |
Reindeer | anti-deer Ig G | KPL Inc. Maryland |
Specific conjugates for IFAT and producer used in game animals.
In a group of game animals, T. gondii antibodies were detected in 32% wild ruminants (50% in sika, 45% red deer, 24% roe deer, 17% fallow deer, 9% mouflon, and in one reindeer), 26% wild boars and 20% hares. The results of serological examination including the number of samples tested, the method and cut-off used, the number and percentage of positive samples, titres obtained in positive samples and reference about published data are summarized in Table 5.
Animals | T. gondii | Assay (cut-off) | Titres | Reference | ||
n | positive | % | ||||
Wild boar | 565 | 148 | 26 | IFAT (40) | 40 – 1280 | Bartova et al. 2006 |
Hares | 333 | 71 | 20 | IFAT (40) | 40 – 640 | Bartova et al. 2010b |
Wild ruminants | ||||||
Red deer | 377 | 169 | 45 | IFAT (40) | 40 – 640 | Bartova et al. 2007 |
Roe deer | 79 | 19 | 24 | IFAT (40) | 40 – 160 | Bartova et al. 2008 |
Sika | 14 | 7 | 50 | IFAT (40) | 80 – 320 | Bartova et al. 2009 |
Fallow deer | 143 | 24 | 17 | IFAT (40) | 40 – 160 | Bartova et al. 2010 |
Mouflon | 105 | 9 | 9 | IFAT (40) | 40 – 320 | Bartova et al. 2011 |
Reindeer | 2 | 1 | IFAT (40) | 80 | Bartova et al. 2012 |
The result of serological examination of game animals with the sample number, the method and cut-off used, titres in positive samples and references.
Experimental studies
In the Czech Republic, two experimental studies were conducted on game animals.
The first study was conducted on hares (Sedlak et al. 2000a). Hares were experimentally infected with T. gondii oocysts. Most infected hares demonstrated behavioural changes, and all of them died between 8 and 19 days. In all hares, parasitemia was demonstrated on days 7 and 12 p.i. T. gondii was isolated from liver, brain, spleen, kidney, lung, heart and skeletal muscles. Based on this result, hares seem to be very sensitive species to T. gondii infection.
The second study was conducted on gallinaceous game birds (Sedlak et al. 2000b). Partridges (Perdix perdix), chukars (Alectoris chukar), wild guineafowl (Numida meleagris) and wild turkeys (Meleagris gallopavo) were inoculated per os with two doses of T. gondii oocysts. Antibodies to T. gondii were detected in the birds by IFAT first on day 7 p.i. Two of five partridges fed 103 oocysts and six of eight partridges fed 105 oocysts died between day 6 and 16 p.i. No clinical symptoms were observed in surviving birds, however enteritis was the most striking lesion in partridges that died. Bioassay in mice revealed T. gondii in the brain, liver, spleen, heart and leg muscles of all partridges and chukars. These results indicate that partridges are highly susceptible to toxoplasmosis, while chukars, wild guineafowls and turkeys seem to be less susceptible.
Wild. boars
In Europe, the highest seroprevalence 100% was found in wild boars from Portugal (Lopes et al. 2011b) or 44% in wild boars from Spain (Closa-Sebastia et al. 2011); while the lowest prevalence 8% was found in wild boars from Slovak Republic (Antolova et al. 2007). In the Czech Republic we found 26% prevalence by IFAT. This prevalence was higher compared to 0% – 15% prevalence found by SFT in the previous studies from the Czech Republic.
The meat of wild boars may harbour tissue cysts of T. gondii and may represent a vehicle of human toxoplasmosis infection. Hejlicek et al. (1997) found tissue cysts in 2% examined wild boars from the Czech Republic, while in the neighbouring Slovakia, T. gondii was isolated from 31% of wild boars (Catar 1972). Hunters and their families consuming meat from wild boars should be aware of T. gondii infection and advised to take precautions. It is highly recommended to cook meat from wild boars thoroughly before human consumption.
Hares
There are several reports of T. gondii infection in hares from Europe (Dubey 2010). The highest seroprevalence 46% was found in hares from Germany (Frolich et al. 2003); in contrast no antibodies were detected in hares from Sweeden (Gustafsson and Uggla 1994). In the Czech Republic, we found 20% prevalence by IFAT. This result is comparable with 4% – 31% prevalence found in previous studies by SFT or MPA. Based on the results of experimental infection, hares seem to be sensitive to T. gondii infection;T. gondii was isolated from liver, brain, spleen, kidney, lung, heart and skeletal muscles (Sedlak et al. 2000).
Wild. ruminants
T. gondii infection in game animals is of epidemiological significance. Deer are strictly herbivores and that is why the high prevalence of T. gondii in deer suggests widespread contamination of the environment with T. gondii oocysts. In red deer, the highest seroprevalence 32% was found by SFT in Scotland (Williamson and Williams 1980), while the lowest 8% by DAT in Norway (Vikoren et al. 2004). We found relatively high prevalence 45% by IFAT in red deer from the Czech Republic. In roe deer, the highest prevalence 63% was found in Norway and Sweden by SFT (Kapperud 1978), while the lowest 13% prevalence was found in Austria by IHA (Edelhofer et al. 1989). In the Czech Republic, we found 24% prevalence by IFAT that was also in range 14% – 58% prevalence found in our country in previous studies. In fallow deer, we found 17% prevalence that is comparable with 24% prevalence found in Spain by MAT (Gauss et al. 2006). In the Czech Republic, we found 9% prevalence in mouflon that is lower compared to 23% prevalence found in France (Aubert et al. 2010). In case of reindeer, only two animals were examined in the Czech Republic. This is very low number that is why it is not possible to compare it with 1% prevalence found in Norway by DAT (Vikoren et al. 20004).
Deer are popular game animals in several countries. The meat of deer may harbour tissue cysts of T. gondii and may represent a vehicle of human toxoplasmosis infection. Toxoplasmosis infection in men was documented after consummation of raw or nearly raw deer meat in USA (Sacks et al. 1983, Ross et al. 2001).
Serological. studies
In a group of zoo animals, 556 animals belonging to 114 species were tested (5 species of primates, 28 species of carnivores, 8 species of perissodactyla and 73 species of artiodactyla). The animals came from 12 zoo and 4 small private exotic centres in the Czech Republic.
Sera of zoo animals were tested for T. gondii antibodies by an IFAT, using the SevatestToxoplasma Antigen IFR (Sevac) and specific conjugates (Table 6). Sera with titer ≥40 were marked as positive.
Order and family | Indirect Fluorescent Antibody Test (IFAT) | |
Conjugate for IFAT | Producer of conjugate | |
Primates | ||
Cercopithecidae | anti-monkey IgG | Sigma-Aldrich s.r.o., Praha |
Hominidae | anti-human IgG | Sevapharma, Praha |
Carnivora | ||
Canidae | anti-dog IgG | Sigma-Aldrich s.r.o., Praha |
Felidae | anti-cat IgG | Sigma-Aldrich s.r.o., Praha |
Hyaenidae | anti-cat IgG | Sigma-Aldrich s.r.o., Praha |
Mustelidae | anti-cat IgG | Sigma-Aldrich s.r.o., Praha |
Otariidae | anti-cat IgG | Sigma-Aldrich s.r.o., Praha |
Ursidae | anti-cat IgG | Sigma-Aldrich s.r.o., Praha |
Viveridae | anti-cat IgG | Sigma-Aldrich s.r.o., Praha |
Perissodactyla | ||
Equidae | anti-horse IgG | VMRD, Pullman, USA |
Artiodactyla | ||
Bovidae | anti-bovine IgG, anti-goat IgG | VMRD, Pullman, USA |
Cameliae | anti-llama IgG | VMRD, Pullman, USA |
Cervidae | anti-deer IgG | KPL, Gaithersburg, Maryland |
Suidae | anti-swine IgG | Sigma-Aldrich s.r.o., Praha |
Specific conjugates for Indirect Fluorescent Antibody Test and their producer used in zoo animals.
In a group of zoo animals, T. gondii antibodies were detected in 193 of 556 (35%) animals, representing 72 of 114 species tested (Sedlak and Bartova, 2006a). According to order, T. gondii antibodies were found in 90% carnivorous, 45% primates, 33% perissodactyles and 22% artiodactyles. According to families, T. gondii antibodies were found in ursidae (100%), felidae (93%), canidae (88%), hominidae (73%), equidae (33%), suidae (29%), cervidae (27%), camelidae (26%), bovidae (20%), cercopithecidae (18%) and in 3 animals of hyeenidae, 2 animals of mustelidae and 2 animals of viveridae. The highest prevalence 100% was found in Eurasian wolf (Canis l. lupus), Maned wolf (Chrysocyon brachyurus) and Sumatran tiger (Panthera t. sumatrae). The highest titre 40960 was found in Pallas´s cat (Otocolobus manul). The results of serological examination of zoo animals are summarized in Table 7.
Order and Family | n | T. gondii | |
Positive | % | ||
Primates | 22 | 10 | 45 |
Cercopithecidae | 11 | 2 | 18 |
Hominidae | 11 | 8 | 73 |
Carnivora | 87 | 78 | 90 |
Canidae | 32 | 28 | 88 |
Felidae | 41 | 38 | 93 |
Hyaenidae | 3 | 3 | |
Mustelidae | 2 | 2 | |
Otariidae | 2 | 0 | |
Ursidae | 5 | 5 | 100 |
Viveridae | 2 | 2 | |
Perissodactyla | |||
Equidae | 46 | 15 | 33 |
Artiodactyla | 401 | 90 | 22 |
Bovidae | 265 | 53 | 20 |
Cameliae | 19 | 5 | 26 |
Cervidae | 110 | 30 | 27 |
Suidae | 7 | 2 | 29 |
The result of serological examination of zoo animals
In the Czech Republic, experimental infection was conducted on budgerigars (Melopsittacus undulatus) that were orally inoculated with T. gondii oocysts with different doses (Kajerova et al. 2003). T. gondii antibodies were found by LAT in all birds. The birds showed no apparent signs of disease. T. gondii was isolated by bioassay in mice from all birds fed 103 or more oocysts. The results show that budgerigars are resistant to T. gondii infection.
Cases of clinical toxoplasmosis in the Czech Republic were recorded in nilgais (Boselaphus tragocamelus) and saiga antelope (Saiga tatarica) (Sedlak et al. 2004). Three captive female nilgais aborted two fetuses and two of their newborn calves died within two days of birth. Parasite T. gondii was demonstrated in the brains and livers of both fetuses and in one of the two neonates by single-stage polymerase chain reaction (PCR) with TGR1E and by semi-nested PCR with B1 gene. Retrospectively, antibodies titers ≥640 were found by IFAT in the sera of all three female nilgais and in one male nilgai used to breed them. Fatal toxoplasmosis was diagnosed in one captive adult female saiga antelope. Tissues cysts of T. gondii were found in the liver, lung, spleen, kidney, and intestine of saiga antelope. Toxoplasmosis was confirmed also by PCR with TGR1E and immunohistochemically. Toxoplasmic hepatitis and pneumonia were considered to be a primary cause of death.
The other cases of fatal toxoplasmosis were recorded in year 2004 in seven Pallas cats in several zoos in the Czech Republic (Sedlak and Vodicka 2005).
There are many reports on toxoplasmosis in zoo animals. Marsupials, New World monkeys, hares and some small ruminants belong to the most sensitive to clinic toxoplasmosis. Fatal toxoplasmosis was also recorded e.g. in captive dik-dik and Pallas cats from zoo in USA (Riemann et al. 1974; Dubey et al. 2002), in lions from a zoo in Africa (Ocholi et al. 1989) and in a Siberian tiger from a zoo in Belgium (Dorny et al. 1989). In the Czech Republic, fatal toxoplasmosis was recorded in saiga and nilgais antelopes from Prague and Chomutov zoos (Sedlak et al. 2004) and in Pallas cats (Sedlak and Vodicka 2005).
In our study, antibodies to T. gondii were found in 90% carnivora, 45% of primates, 33% perissodactyla and 22% artiodactyla. When compared to other similar study concerning zoo animals, T. gondii antibodies were found in 47% carnivora, 25% artiodactyla and 23% primates (Gorman et al. 1986). We found 93% prevalence in felids; that is higher when compared with 32%, 64.9% or 75.8% prevalence found in felids from zoo in California (Riemann et al. 1974), Brazil (Silva et al. 2001) and Florida (Lappin et al. 1991), respectively.
The potential source of T. gondii infection for carnivores is meat contaminated with T. gondii tissue cysts; herbivores can be infected by food contaminated with T. gondii oocysts and omnivorous animals by both ways. To prevent spreading of T. gondii infection among zoo animals, cats, including all wild felids should be housed in buildings separated from other animals, particularly the most sensitive marsupials and New World monkeys. There must be protection against free access of domestic cats to sources of food and water or into the buildings with animals, especially those that are the most sensitive to toxoplasmosis. Feline faeces should be removed daily to prevent sporulation of oocysts.
Further work should focus on serological studies in other animal groups that are neglected but may represent a risk of infection for humans in case of consumption of their meat or other products. Such animals include, for example, rabbits, ostriches, pigeons, pheasants and mallard ducks. In addition, rodents, wild birds and wild carnivores (foxes, marten and others) may play an important part in the circulation of T. gondii infection in nature and thus represent a risk of infection for wildlife, domestic animals and human people alike. Serological studies should be supplemented with an evaluation of the infection risk factors and with the use of molecular methods to detect T. gondii in animal products, as well as to characterize T. gondii genotypes circulating in animal populations in the Czech Republic.
The results obtained in last 10 years were supported by the Ministry of Education, Youth and Sports of the Czech Republic (Grant No. MSM6215712402). We would like thank to R. Vodička, J. Váhala and F. Treml for their assistance in the collection of the serum samples and thank to students (V. Říhová, Z. Satková, H. Michnová, M. Syrová, M. Šíblová, H. Říhová, A. Šedivá, J. Drastíková) for their assistance with serological examinations.
Increase in global temperature had major impact on crop productivity especially in tropical and sub tropical regimes. Based on climate model predictions, around 1.8–4.0°C rise in air temperature was expected in 21st century [1]. The increase in temperature beyond a certain threshold level tends to induce detrimental effects in plant growth and development. In general, the elevation in temperature of 10–15°C above ambient triggers heat shock in crop plants. The extent of induced heat stress depends on the duration, intensity and rate of increase in global air temperature [2]. Indian lowlands share 15 per cent of global wheat production. The change in global climate would shift these fertile lowlands into heat stressed unproductive environment [3]. Similarly, the cultivation of cereals in Southern Africa and South East Asia was predicted to be heat stressed zone in near future [4]. Around 4–14% yield decline in rice was encountered due to elevated temperature of 1°C in South-East Asia [5]. The declined productivity due to elevated temperature imposes the urgent need for development of climate resilience genotypes. Evolving heat tolerant cultivars would highly benefit the livelihood of developing countries as around 70–80% of population relies on agriculture. Understanding the effect of heat stress on crop plants and its adaptation mechanisms would help in framing out the breeding strategies for high temperature tolerance.
\nHeat tolerance in crop plants is a complex mechanism involving adaptations through altered physiological process, morpho-anatomical features and induction of several biochemical pathways. On exposure to high temperature, several signal transduction pathways were triggered leading to changes in gene expression. As a result, varied stress related proteins were synthesized contributing heat tolerance in plants [6]. The tolerance mechanism to high temperature stress varies within genotypes of a plant species. The existing variation between and within species provide scope for evolving heat tolerant lines through conventional breeding approaches [7]. Dissecting out genetic information through molecular tools would hasten the development of climate resilient cultivars contributing to food security in near future. A brief review on plant response, adaptation mechanisms and genetic approaches to combat heat stress were presented in this chapter.
\nHeat stress had varying impact on different phenological stages viz., germination, seedling, vegetative, flowering and reproductive of crop plants [8]. The plant response to heat stress depends on the duration, degree of rise in temperature and plant type. Under tropical regimes, high temperature with intense solar radiation poses a major limiting factor for yield by inducing leaf abscission, leaf senescence, scorching of leaves, branches and stems, growth inhibition, pollen infertility and poor seed formation [9, 10]. A significant decline in relative growth rate, shoot dry weight and net assimilation rate was recorded in sugarcane, maize and pearl millet on exposure to high temperature stress [11]. High reduction in grain quality was recorded in most of the cereal crops grown under heat stress environments [12]. Several physiological processes such as partitioning of assimilates, plant-water relations and shoot growth was affected due to heat stress in common bean [13]. In general, the susceptibility to heat stress was found higher at reproductive stage of plant development. An excessive yield loss is recorded in legumes on exposure to high temperature (30–35°C) during anthesis stage [14]. Drastic reduction in grain number and weight was observed in wheat at high temperature regimes [15]. Heat stress affects several metabolic pathways leading to accumulation of reactive oxygen species (ROS) which is a major component for oxidative stress in crop plants [16]. The photosystem centres (PS I and PS II) of chloroplast, mitochondria and peroxisomes are the major sites for generation of ROS in plants [17]. High temperature stress disrupts the stability of cell membrane through protein denaturation [18]. The induction of ROS due to high temperature stress was correlated with premature leaf senescence in Gossypium sp. [19]. Accumulation of ROS in root cells was evidenced in wheat on exposure to high temperature for two days [20].
\nPlants tend to adapt several complex mechanisms through phenological and morphological changes to combat high temperature stress (Figure 1). On heat stress regimes, plants exhibit varied short term escape/avoidance mechanisms viz., altered leaf orientation, transpirational cooling, altered membrane lipid properties, early maturation and so on for its survival. Plants show varied degree of leaf rolling upon intensity of solar radiation. A significant tolerance to high temperature was observed in wheat by maintenance of water potential in flag leaf through adoption of leaf rolling under heat shock conditions [21]. Increase in trichomatous and stomatal densities, waxy layer on leaves, and larger xylem vessels are the common features induced during heat stress [22]. On contrary, plants also evolve long term tolerance mechanisms for its effective survival and productivity under high temperature. Induction of osmoprotectants, antioxidants, late embryogenesis abundant proteins, dehydrins, and heat shock proteins are the major factors involved in counteracting the heat shocks. Accumulation of osmolytes such as proline, trehalose, and glycine betaine plays a vital role in imparting tolerance via cellular osmotic adjustment, detoxification of ROS, stabilization of enzymes and membrane proteins [23]. Several enzymatic and non-enzymatic antioxidant defense components are also involved in protection against oxidative stress induced by free radicals [24]. The activities of ROS scavenging enzymes are temperature specific. In general, most of the antioxidant enzymes show increased activity with elevation in temperatures. It is also influenced by genotype, growing season and phenological stages of plant [25]. Under high temperature conditions, several signaling molecules such as nitrous oxide, Ca-dependent protein kinases, Mitogen mediated protein kinase, sugars, and phytohormones play a role in stimulation of stress responsive genes via transduction pathways [26]. Evolving adaptation mechanisms (either tolerance or avoidance) to high temperature and drought would be more rewarding at arid conditions as it is often correlated.
\nAdaptation mechanisms for high temperature tolerance in crop plants.
Breeding for high temperature tolerance requires an essential knowledge on plant adaptation response to heat shocks. In general, the genotypes exhibiting less detrimental effect on photosynthesis and reproductive development tend to survive well under heat prone areas [27]. Involvement of these two components in selection criteria would be beneficial in evolving thermo tolerant cultivars. Tolerant genotypes evolve several morphological, physiological and biochemical alterations in response to heat shocks. Knowledge on sensitivity of several phenological stages to high temperature will pave way for trait specific improvement. High temperature is often correlated with other environmental factors which poses a major limitation for selection under field conditions. At present, varied selection criteria has been developed by scientists, which favors delineation of superior variety at prevailing environment [28]. Heat tolerant index has been evolved for sorghum which depicts the proportion of growth recovery after exposure to high temperature stress. It is the ratio of increase in coleoptile growth in a heat stress environment [50°C] to the enhancement in coleoptile length under normal environment (non-stress) [29]. It proves cost effective and rapid method to screen a large population size within shorter period. A proper validation of such technique would facilitate the development of tolerant lines in other crop species. Pollen viability and fruit set was considered as major selection criteria to predict yield under high temperature stress in tomato [30]. Physiological based trait selection such as harvest index, photosynthetic efficiency, respiration rate, delayed senescence and canopy architecture will also contribute towards increased tolerance to heat stress [31, 32].
\nInter-mating among closely related individuals for improvement of economic traits resulted in decline of genetic variability in a crop species [33]. Characterization of gene pool including land races and wild relatives would offer several tolerant genes for abiotic tolerance. Extensive efforts were made in screening of heat tolerant genotypes which can be directly introduced as a cultivar or utilized to introgress gene into new genetic background [34]. Thermo-tolerant lines were successfully isolated from wild gene pool in wheat [35]. High magnitude of variation was observed in wild progenitor “Aegilops tauschii” of wheat for cell viability and membrane stability [36]. Similarly, a heat tolerant source for reproductive stage was identified in A. geniculata and A. speltoides Tausch which would pave way in development of thermo-tolerant hexaploid wheat cultivars in near future [37]. A higher growth rate and improved photosynthetic efficiency was observed in wild relative “Oryza meridionalis” of rice at high temperature [38]. Indirect selection on pollen viability led to identification of thermo-tolerant accessions in soybean (DG 5630RR) [39], chickpea (ICC15614 & ICC1205) [40], maize (AZ100) [41], and several other crop species. Direct selection based on yield under target environment (heat stress) resulted in development of tolerant lines in many tropical grain legumes. Four tolerant genotypes/accessions viz., SRC-1-12-1-48, SRC-1-12-1-182, 98012-3-1-2-1 and 98020-3-1-7-2 were isolated in common bean by employing stress tolerant indices [42]. Nine thermo-tolerant wild accessions were delineated in USDA upland cotton germplasm by employing chlorophyll fluorescence technique [43].
\nEvolving thermo-tolerance through conventional breeding approach proves promising in many crop species. Breeding for early maturing genotype in broccoli had improved head quality by avoiding heat stress at flowering stage [44]. In general, breeding programmes are carried out in hotter regions which promote selection of thermo-tolerant traits. Physiological based trait breeding was practiced at International Maize and Wheat Improvement Center (CIMMYT) for development of heat tolerant cultivars in wheat. The parental genotypes were characterized through various crossing schemes and appropriate breeding programme was framed for improvement of thermo related traits [45]. A wild ancestor “T. tauschii” was utilized as a gene donor for achieving increased grain size and filling percent under high temperature through recurrent selection [46]. Similarly, three cycles of recurrent selection had led to improved yield under heat stress regimes in potato [47]. Thermo tolerant alleles were introgressed into heat sensitive cultivar “Paymaster 404” from a donor accession “7456” of G. barbadense through backcross breeding [48]. A significant improvement in yield was realized under heat stress environment by adoption of gametic selection in maize [41]. A deep rooted cultivar “Nagina 22 (N22)” of aus rice exhibited high pollen viability and spikelet fertility (64–86%) under heat stress [49]. The thermo-tolerance of N22 was successfully introgressed into Xieqingzao B line through backcross method [50]. Dissecting out the genetic and physiological basis of thermo-tolerance will hasten up the development of resilient cultivars suited to hotter regions.
\nThe genetic basis of thermo-tolerance is not clearly understood because of complex trait inheritance. Advances in molecular approaches such as DNA marker identification and genotyping assay had paved way in determination of several QTL’s associated with high temperature tolerance [51]. In wheat, QTL’s were identified for canopy temperature, and chlorophyll fluorescence imparting tolerance to heat stress [52]. A major QTL “Htg 6.1” in lettuce was involved in enhancement of seed germination capacity at high temperature [53]. A recessive QTL for increased spikelet fertility under high temperature was dissected out in rice at chromosome 4. The identified QTL were found in several populations of heat tolerant rice cultivars [54]. Six QTL’s were involved to enhance fruit set at high temperature in tomato [55]. Five thermo tolerant QTL’s were identified in Brassica campestris by employing random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers [56]. In maize, eleven major QTL’s for increased pollen germination and pollen tube growth under high temperature was mapped using restriction fragment length polymorphism (RFLP) markers [57]. Identification of candidate QTL’s would pave way in precise introgression of heat tolerant genes into superior cultivars through marker assisted breeding approach.
\nThe closely associated markers with targeted QTL will hasten the recovery of superior genotypes with heat tolerant traits in a population. A marker assisted breeding approach was employed in rice to derive heat tolerant line with superior grain quality. Two flanking markers viz., ktIndel001 and RFT1 enclosing 1.5 Mb chromosomal region was transferred from tolerant cultivar “Kokoromachi” to Tohoku 168. Significant improvement in grain quality under high temperature was observed in the derived NIL’s compared to susceptible cultivar “Tohoku 168” [58]. Fourteen SSR markers linked to heat susceptibility index of grain filling per cent and single kernel weight was identified in bread wheat which was employed in marker assisted selection (MAS) to screen genotypes for thermo tolerance [59]. Utilization of MAS approach for heat tolerance remains less efficient because of high gene x environment and epistatic interactions. The low breeding efficiency can be resolved by genomic selection (GS) approach which involves wide number of molecular markers exhibiting high genome coverage. High genetic gain is realized in GS approach due to close association between predicted and true breeding value over generations [60].
\nAt present, transgenic approach also proves to be desirable tool for designing thermo tolerant lines via introgression of genes from diverse gene pools [61]. The genetic transformation was focused primarily on transcription factors, induction of heat shock proteins, molecular chaperones, osmolytes, antioxidant components and growth regulators [62]. Heat shock proteins play a primary role in imparting thermo tolerance in crop species. It is functionally associated with diverse group of molecular chaperones that is involved in restoration of degraded proteins to their native structure under high temperature. Induction of heat shock proteins through genetic manipulation was achieved in arabidopsis [63], maize [64], rice [65], soybean [66], and pepper [67]. The DREB gene family was also reported to impart heat tolerant response in many crop species. Over expression of ZmDREB2A in maize [68] and GmDREB2A in soybean [69] was associated with increased survival and adaptation under high temperature. Transgenic techniques were employed to alter membrane lipid properties for thermo-tolerance in crop species. High proportion of saturated fatty acid in membrane had increased tolerance under heat stress. Suppression of omega-3 fatty acid desaturase gene in chloroplast had reduced the accumulation of trieonic fatty acid in transgenic tobacco [70] and tomato [71] leading to thermo-tolerance. A significant accumulation of glycine betaine (osmolyte) was achieved in arabidopsis through transfer of “cod gene” from Arthrobacter globiformis [72]. High proportion of glycine betaine protects the PSII component by inhibiting the ROS activities under heat stress. Implementation of transgenic approaches in other crop species will accelerate the development of resilient genotypes suited to high temperature regimes.
\nDevelopment of thermo-tolerant lines has to be prioritized to meet out the future climatic change coupled with food demands. Knowledge on plant response and adaptation mechanisms to heat stress is required for framing out breeding strategies. It remains a challenging task in evolving resilient genotypes suited to high temperature because of less efficient screening protocols at field conditions. The existence of low genetic variation for heat response related traits limited the progress of conventional breeding approach in many crop species. Use of molecular breeding strategies had opened up several heat tolerant related QTL’s in crop species. However, still precise research work involving huge marker data is needed for attaining high breeding efficiency for thermo tolerance. Recently, the involvement of transgenic approach paved way for utilization of tolerant source from diverse gene pools. Study on induction of heat shock proteins led to increased thermo tolerance in many crop species. Similarly, other heat response related traits such as induction of antioxidant components, osmolytes, and chaperones were also included in transgenic approach for inducing heat stress tolerance. Thus, high economic yield could be realized at elevated temperature regimes with the involvement of combined breeding approaches.
\nThe authors are highly thankful to Dr. V. Geethalakshmi, Director, Directorate of Crop Management, Tamil Nadu Agricultural University (TNAU) for her valuable suggestions towards this chapter. We also acknowledge Dr. P. Jayamani, Professor and Head, Department of Pulses, TNAU; Dr. M. Raveendran, Professor and Head, Department of Biotechnology, TNAU; and Dr. K. Ganesamurthy, Professor and Head, Department of Rice, TNAU for rendering supportive documents on high temperature tolerance.
\nThe authors declare no conflict of interest towards this chapter.
The authors express their gratitude to the Directorate of Crop Management for providing scientific support on high temperature tolerance.
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