Biological ability of Malpura rams to counter heat stress challenges and its consequences on production performance in a semi-arid tropical environment

A study was conducted to assess the biological ability of Malpura rams to counter heat stress challenges and its impact on the productive performance in a semi-arid tropical environment. The eighteen adult Malpura rams (average body weight 55.0 kg) were divided into two groups, GI (n = 9; Control) and GII (n = 9; heat stress). The study was conducted for a period of 60 days. The results showed that body weight, body condition score, heart girth and feed intake were significantly (p < 0.05) lower in GII as compared to GI rams. The physiological responses i.e. respiration rate, pulse rate, rectal temperature and sweating rate (body and scrotum) were significantly (p < 0.05) higher in rams exposed to heat stress. Among the endocrine parameters, plasma T₃, T₄ and testosterone concentrations were significantly (p < 0.05) lower in GII rams, while cortisol concentration was significantly (p < 0.05) higher in GII rams. In addition, heat stress significantly (p < 0.05) reduced semen volume, total sperm motility, sperm concentration and progressive sperm motility in GII rams. The results indicated that Malpura rams adapted to the heat stress condition, however, while doing so their production performances were compromised.     

Effect of walking stress on growth,physiological adaptability and endocrine responses in Malpura ewes in a semi-arid tropical environment

Sheep in hot semi-arid environments are mostly reared using extensive systems. In addition to thermal stress and feed scarcity, the animals need to walk long distances for grazing in this ecological zone. A study was conducted to assess the effect of long-distance walking on adaptive capability in terms of physiological, biochemical and endocrine responses in Malpura ewes. Fourteen adult Malpura non-pregnant ewes weighing between 33 and 35 kg were used in the study. The ewes were randomly allocated into two groups of seven animals each: GI (n = 7; Control), and GII (n = 7; walking stress). The animals were stall-fed with a diet consisting of 70% roughage and 30% concentrate. Both GI and GII ewes had uniform access to feed and water. The walking stress group (GII) ewes were made to walk 14 km in two spans between 0900 and 1500 hours with 1 h 30 min for each span (7 km) of walking. The ewes subjected to walking stress (GII) were prevented from grazing by applying a face mask made of cotton thread. The study was conducted for a period of two estrous cycles (35 days) during the autumn season (October–November). Physiological responses were recorded twice daily at 0800 and 1400 hours at weekly intervals. Blood samples were collected from the jugular vein at weekly intervals to study the effects of walking stress on blood biochemical and endocrine parameters. The results indicate that walking stress had significant (P < 0.05) influence on body weight, average daily gain, respiration rate (RR), rectal temperature (RT), haemoglobin (Hb), packed cell volume (PCV), plasma glucose, calcium, phosphorus, aspartate amino transferase (AST), alanine amino transferase (ALT), tri-iodo-thyronine (T₃), thyroxin (T₄), and cortisol. However, walking stress did not influence the reproductive hormone levels. The significant changes in RR, RT, plasma cortisol, T₃ and T₄ show that Malpura ewes have the capability to adapt to long-distance walking, and that adrenal and thyroid gland hormones play a significant role in such adaptation.     

Adaptive capability as indicated by endocrine and biochemical responses of Malpura ewes subjected to combined stresses (thermal and nutritional) in a semi-arid tropical environment

A study was conducted to assess the effect of combined stresses (thermal and nutritional) on endocrine and biochemical responses in Malpura ewes. Twenty eight adult Malpura ewes (average body weight 33.56 kg) were used in the present study. The ewes were divided into four groups viz., GI (n = 7; control), GII (n = 7; thermal stress), GIII (n = 7; nutritional stress) and GIV (n = 7; combined stress). The animals were stall fed with a diet consisting of 60% roughage and 40% concentrate. GI and GII ewes were provided with ad libitum feeding while GIII and GIV ewes were provided with restricted feed (30% intake of GI ewes) to induce nutritional stress. GII and GIV ewes were kept in climatic chamber at 40°C and 55% RH for 6 h a day between 1000 hours and 1600 hours to induce thermal stress. The study was conducted for a period of two estrus cycles. The parameters studied were Hb, PCV, glucose, total protein, total cholesterol, ACP, ALP, cortisol, T₄, T₃, and insulin. Combined stress significantly (P < 0.05) affected all parameters studied. Furthermore, the results revealed that, compared to thermal stress, nutritional stress had a less significant effect on the parameters studied. However, when both these stresses were coupled, they had a severe impact on all the parameters studied in these ewes. It can be concluded from this study that two stressors occurring simultaneously may impact severely on the biological functions necessary to maintain homeostasis in sheep.     

Effect of thermal exposure on physiological adaptability and seminal attributes of rams under semi-arid environment

Thermal stress in hot semi-arid environment is a major limitation of sheep production in tropical and sub-tropical climatic condition. The animals tend to maintain homeostasis through physiological adjustments in a hot environment (maximum temperature reaches up to 47.5 °C). Therefore, the present study was carried out to assess the effect of thermal exposure on physiological adaptability and seminal attributes of rams under semi-arid environment. The experiment was conducted for eight weeks involving sixteen Malpura crossbred rams (GMM: Garole X Malpura X Malpura). The rams were divided equally into two groups, designated as G1 and G2, respectively. The rams in G1 (Control) group were kept in a sheep shed under naturally prevailing environment without artificial manipulation of ambient temperature (Temperature 30.48±0.38 °C; Relative Humidity 28.59±1.15%). The rams of G2 group were exposed to different temperature at different hours of the day (38 °C at 1000–1100 h; 40 °C at 1100–1200 h; 42 °C at 12:00–1300 h; 43 °C at 1300–1400 h; 44 °C at 1400–1500 h and 42 °C at 1500–1600 h) in a climatic chamber for thermal exposure. Physiological responses, blood biochemical profile, blood endocrine profile, sexual behavior and seminal attributes were measured for both the groups. Thermal exposure significantly (P<0.05) increased the water intake; respiration rate, rectal temperature and skin temperature at afternoon in rams. Exposure of rams to thermal stress (G2) significantly (P<0.05) increased cortisol level and decreased tri-ido-thyronine level. The latency period after the first ejaculation, decreased significantly (P<0.05) in G2. The percentage of rapid motile sperm, linearity and average path velocity of sperm were also altered significantly (P<0.05) in thermal exposed rams as compared to control. However, comparable feed intake, body weight, and major blood biochemical parameters, as well as acceptable semen quality attributes of all the rams indicated that the Fec B gene introgressed Malpura cross rams adapted to the thermal exposure under semi-arid tropical climate.     

Effect of multiple stress factors (thermal,nutritional and pregnancy type) on adaptive capability of native ewes under semi-arid environment

This study was conducted to evaluate the effect of multiple stress factors (thermal, nutritional and pregnancy type) on two different native track breeds of ewes as reflected by their adaptive capability under semi-arid environment. The multiple stressor experiment was conducted in twenty-four ewes (12 Santa Inês and 12 Morada Nova ewes). Both heat stress and pregnancy stress was common to all four groups. However, the animals were divided into further two groups within each breed on the basis of nutrition regimen. According the groupings were: Group 1 (Six Santa Ines ewes; heat stress; nutrition at 0.5% of BW; single pregnancy); Group 2 (Six Santa Ines ewes; heat stress; nutrition at 1.5% BW; twin pregnancy); groups Group 3 (Six Morada Nova ewes; heat stress; nutrition at 0.5% of BW; single pregnancy); Group 4 (Six Morada Nova ewes; heat stress; nutrition at 1.5% BW; twin pregnancy). All the animals in the experiment were pregnant. Heat stress was induced by exposing all animals to summer heat stress in outside environment while the nutritional regimen followed was at 0.5% and 1.5% level of body weight (BW) respectively in each breed. The experiment was conducted in a completely randomized design with two breeds, two nutritional treatments and two pregnancy types, 10 repetitions for physiological parameters and six for blood parameters, with repeated measures over time. Physiological parameters (respiratory rate, pulse rate and rectal temperature) were measured with the animals at rest in the morning and afternoon, 0600-0700 and 1300–1400 h, respectively, every seven days. Blood samples were collected every 14 d for determination of serum glucose, triglycerides, cholesterol, urea and creatinine. We found interaction effect between breed and pregnancy type on respiratory rate and rectal temperature with greater values in Santa Inês ewes than Morada Nova ewes. However, there was no significant fixed effect of pregnancy type and supplementation level on physiological responses of breeds. Environmental factor (period of the day) had influenced the physiological responses of ewes during all gestational period. Santa Inês ewes had greater serum glucose concentration at 105 d and 120 d of gestation compared to the Morada Nova ewes. Morada Nova ewes had greater concentrations of triglycerides, urea at 120 d, 150 d and also greater cholesterol at 105 d, 135 d and 150 d of gestation compared with Santa Inês ewes. The present result indicates that thermal condition was most important factor that modified the physiological responses of ewes in a semi-arid tropical environment.     

Evaluation of gross anatomical features of cervix of tropical sheep using cervical silicone moulds

The lambing rate obtained following cervical artificial insemination (AI) with frozen semen in sheep is low mainly due to the inability of frozen-thawed sperm to traverse the tortuous nature of the cervical canal. Although acceptable fertility has been attained by circumventing the cervical barrier through laparoscope aided intrauterine AI, the emphasis is currently given on the development of alternate non-invasive transcervical AI procedures. The complex anatomy of the cervix does not facilitate easy transcervical passage for an insemination catheter. The aim of the present study was: (i) to examine the gross anatomy of the cervix in slaughtered ewe lambs and adult ewes of the native Malpura and Kheri breeds raised under semi-arid tropical environment; and (ii) to cast silicone moulds of the reproductive tracts for measuring the dimensions of the cervix. Eighty reproductive tracts were excised immediately from carcass of Malpura and Kheri ewes and the external os of each one was classified depending on their appearance as duckbill, spiral, rosette or flap. The cervical canal of each tract was filled with a silicone sealant for casting the mould. Fifty complete silicone moulds were obtained representing 25 from ewe lambs and 25 from adult ewes. The mean lengths of the cervical mould of ewe lambs and adult ewes were 3.8+/-0.12 and 5.3+/-0.15 cm, respectively. The average number of funnel shaped folds in the cervical mould of ewe lambs and adult ewes were 3.2+/-0.19 and 3.4+/-0.22. However, the second and third-folds from the os were observed to be accentric in both ewe lambs and adult ewes. The information generated in this study would be useful for increasing the success rate of penetration in ewes exhibiting estrus in order to improve the lambing rate of tropical ewes following transcervical AI.     

Comparative physiological responses of stress in sheep selected for resistance or susceptibility to Haemonchus contortus in semi-arid tropical region during different seasons

The present study was conducted to assess the comparative physiological responses of stress in sheep selected for resistance or susceptibility to Haemonchus contortus in the semi-arid region during different seasons. A total of 20 adult Malpura sheep from resistant and susceptibility line were selected for study. The intensity of strongyle infection, blood biochemical and cortisol levels were estimated monthly basis while meteorological data were collected everyday throughout the year. The temperature-humidity index was the highest during dry-hot season. The faecal egg count increased significantly (p < 0.05) during June–August (rainfall season) in susceptible sheep as compared to resistant sheep. The plasma cortisol level increased significantly (p < 0.05) in both the groups during dry hot (March–May) season and it remained higher in the susceptible line till June–August. The total plasma protein reduced significantly during hot-humid with scantly rainfall (September–November) season as compared to other season in susceptible line. It is pertinent to conclude that though sheep remained in stress during dry hot (March–May) season, but effects of stress on the intensity of strongyle infection and blood biochemical were better tolerated by animals of resistant line during hot-humid season compared to susceptible line under semi-arid region.     

Sperm motion characteristics of GarolexMalpura sheep evolved in a semi-arid tropical environment through introgression of FecB gene

The arid and semi-arid tropical climates of India are endowed with vast diversity of non-prolific sheep breeds. The GarolexMalpura sheep has been evolved in a semi-arid tropical environment by introgression of FecB gene via artificial insemination of Malpura ewes using diluted semen of prolific microsheep Garole and subsequently multiplied by inter se mating among GarolexMalpura halfbreds. The aim of the present study was to identify FecB mutation in sexually mature GarolexMalpura rams by forced RFLP-PCR of BMPR-1B gene and evaluate: (i) semen production and sperm motion characteristics of GM rams and (ii) influence of age and FecB genotype on their semen attributes. Semen was collected during autumn season from 12 donor rams by artificial vagina on 8 occasions at weekly interval. The overall means of traits which did not differed significantly with age or FecB genotyping were volume (0.72 ml), mass motility (4.44), sperm concentration (2721.56 x 10(6)ml(-1)), curvilinear velocity (134.51 microm/s), motility (81.3%), amplitude of lateral head displacement (6.24 microm), beat frequency (44.43 Hz), sperm head elongation (48.9%) and sperm head area (10.01 microm(2)). The FecB genotyping had a significant effect (P<0.05) on percent linearity and rapid motile sperms, which did not vary significantly with age. Although sperm concentration was higher in FecB(BB) and FecB(B+), compared to FecB(++) genotypes but the effect was non-significant. The age and FecB genotyping had significant effects (P<0.05) on straightness, average path velocity, straight-line velocity and percentage of medium or slow motile sperms. It is concluded that GarolexMalpura rams with introgressed FecB gene are capable of producing good quality semen in a semi-arid tropical climate.     

Biological functions as affected by summer season-related multiple environmental stressors (heat,nutritional and walking stress) in Malpura rams under semi-arid tropical environment

The study was conducted with the primary objective to establish the impact of simultaneously imposed multiple stressors (thermal, nutritional and walking) on various biological functions in Malpura rams. The study was conducted for a period of 45 days. Twenty adult Malpura rams (average BW 44.9 Kg) were used in the present study. The rams were divided into two groups, CON (n = 10; Control) and MS (n = 10; multiple stressors). Both body weight (p < 0.05) and body condition scoring (p < 0.01) were lower in MS compared with CON rams. All the scrotal measurements reduced significantly (p < 0.01) in MS rams. All the testicular measurements also reduced significantly (p < 0.01) in MS as compared to CON rams. The seminal attributes, individual motility (p < 0.05), rapid motility (p < 0.05) and curvilinear velocity (p < 0.05) were significantly reduced in CON rams. However, slowness (p < 0.05), linearity (p < 0.05) and straightness (p < 0.05) parameters significantly increased in MS as compared to CON rams. In addition, plasma testosterone concentration was lower (p < 0.05) in MS rams. The study established the impact of multiple stressors on growth and reproductive performance of Malpura rams. Further, the results revealed that the only semen quality was affected by multiple stressors, but the semen production remained intact between the groups, indicating the extreme adaptive capability of Malpura rams to hot semi-arid tropical environment.     

Improving productivity of Malpura breed by crossbreeding with prolific Garole sheep in India

Garole, a prolific small sized sheep breed of India was introduced in the Mutton Project of the Institute during 1997 to explore the possibility of incorporating fecundity gene into Malpura (M) sheep of semi-arid region of Rajasthan. The data on reproduction and production traits of M and Garole × Malpura (GM) half-breds was used in the study to assess the improvement in evolved crossbred stock. The overall percent gain in prolificacy in GM was 52.38, which increased to 75.73 in third parity. The GM compared to M ewes also weaned more lambs per ewe lambed (1 versus 1.41). The lambing rate averaged 91.75 and 148.21% in M and GM ewes, respectively. The twin lambing percentage was only 4.71% in M flock while; it reached up to 51.10% in GM. Moreover, 5.72% of GM ewes produced triplets also. The genetic group non-significantly affect ewe efficiency (EE), but GM produced 3.3% more total lambs weight per ewe lambed at weaning and 6.15% more total lamb weight per ewe lambed at 6-month of age. The body weights at different ages of GM half-breds were lower (P < 0.01) compared to contemporary M lambs. The parity of ewes significantly affects body weight from birth weight to 12-month of age. The survivability of GM half-breds was almost at par with the local M sheep. Results revealed that the fecundity genes responsible for increasing prolificacy have been incorporated into the GM and it might prove a valuable germplasm towards evolving a new prolific strain of sheep. The crossing of Garole with M increased the ewe efficiency.     

Effects on fetal and maternal body temperatures of exposure of pregnant ewes to heat, cold, and exercise.

We exposed Dorper-cross ewes at approximately 120-135 days of gestation to a hot (40 degrees C, 60% relative humidity) and a cold (4 degrees C, 90% relative humidity) environment and to treadmill exercise (2.1 km/h, 5 degrees gradient) and measured fetal lamb and ewe body temperatures using previously implanted abdominal radiotelemeters. When ewes were exposed to 2 h of heat or 30 min of exercise, body temperature rose less in the fetus than in the mother, such that the difference between fetal and maternal body temperature, on average 0.6 degrees C before the thermal stress, fell significantly by 0.54 +/- 0.06 degrees C (SE, n = 8) during heat exposure and by 0.21 +/- 0.08 degrees C (n = 7) during exercise. During 6 h of maternal exposure to cold, temperature fell significantly less in the fetus than in the ewe, and the difference between fetal and maternal body temperature rose to 1.16 +/- 0.26 degrees C (n = 9). Thermoregulatory strategies used by the pregnant ewe for thermoregulation during heat or cold exposure appear to protect the fetus from changes in its thermal environment.     

Inability of Ceratitis capitata (Diptera: Tephritidae) to overwinter in the Judean hills

The overwintering potential of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), in cold winter areas within its northern distribution is a key element in understanding its ecology. Recent studies have suggested that although originating in tropical Africa, the fly has become adapted to the cold weather that prevails within its northernmost areas of distribution. We address the question of whether the Mediterranean fruit fly has expanded its overwintering range to include the mountains of central Israel. Doing so would imply that the fly has developed either a behavioral or a physiological mechanism to cope with low temperature and/or damp conditions in combination with cold. We monitored adult populations year round, sampling fruit, calculating expected emergence days for overwintering flies, and studying adults captured within dense and sparse apple orchards. We also performed several manipulative experiments to study preimago ability to survive the winter under natural or seminatural conditions. The study was conducted in the central mountains of Israel at 700-m altitude from 1994 to 2003. Comparison experiments also were conducted at 400 m and at sea level. Our results show 1) no adults captured during the winter and spring, 2) an absence of new infestations during the winter and spring, and 3) inability of preimago stages to overwinter in the central mountains of Israel. Thus, we conclude that the fly does not overwinter in the central mountains of Israel. We discuss the ecological and applied significance of our findings.     

Seasonal variations in physio-biochemical profiles of Indian goats in the paradigm of hot and cold climate

To explore the heat and cold adaptation in Indian goats by the physiological, haematological, blood biochemical parameters and their seasonal variations, this study was conducted on heat- and cold-adapted Indian goats maintained in their natural habitat. Study was carried out in three different phases coincide with the three seasons (winter, spring and summer). The levels of physiological responses, that is rectal temperature, respiration rate and pulse rate, were observed to be significantly (p < 0.01) lower in heat-adapted breeds and higher in cold-adapted breeds, whereas the levels of Hb, PCV and TEC were significantly (p < 0.01) higher in cold-adapted goats. Significantly (p < 0.01) higher levels of plasma thyroid hormones (thyroxine and triiodothyronine) and plasma stress enzyme (AST and ALT) were also observed in cold-adapted goats. Significant (p < 0.01) seasonal variations in physiological responses, haematological and blood biochemical parameters in both heat- and cold-adapted breeds were reported in this study. Physiological responses, plasma enzymes and plasma cortisol levels significantly (p < 0.01) increased during summer in all the goat breeds. The levels of haematological parameters (Hb, PCV, TEC and TLC) and plasma thyroid hormones (thyroxine and triiodothyronine) decreased during summer. The changes in physiological parameters during summer due to heat stress were higher in cold-adapted goats whereas the levels of changes in these parameters during winter due to cold were higher in heat-adapted goats. High neutrophil/lymphocyte ratio during summer in cold-adapted breeds is an indicator of higher level of stress. Decrease in plasma electrolytes (Na and K) during summer also observed in cold-adapted breeds during summer. The variations in physiological, haematological, blood biochemical parameters in heat- and cold-adapted goats may be due to their adaptation to different environmental and geographical conditions essential for their survival.     

Studies on effect of Booroola (FecB) genotype on lifetime ewes' productivity efficiency, litter size and number of weaned lambs in Garole x Malpura sheep

The FecB gene of Garole was introgressed into non-prolific Malpura sheep. The present study was conducted to evaluate the effects of FecB genotypes on cumulative lifetime (three lamb crops) litter size (CLS), cumulative number of weaned lambs (CWL) and cumulative ewe's productivity efficiency (CEPE) in 51 Garole x Malpura (GM) crossbred sheep. The GM ewes of F(1) were selected and screened for FecB mutation using forced RFLP-PCR technique. The majority (78.4%) of F(1) GM individuals were carriers (FecB(B+)) for the FecB mutation. In first parity 55% FecB(B+) ewes gave births to multiple lambs. The FecB genotypes were significantly (P<0.01) associated with the CLS and CWL. The FecB(B+) ewes resulted in 65.6 and 62.1% higher CLS and CWL, respectively compared to non-carriers. The CEPE was also affected significantly by the FecB genotypes at birth, weaning, 6 and 12 months of age. The FecB(B+) ewes weaned 20.9% higher total litter weight as compared to FecB++ ewes and at 12 months age the difference was 43.5% in favor of B+ ewes. The study indicated that the CLS, CWL and CEPE of carrier ewes (FecB(B+)) were comparatively higher than that of non-carriers (FecB++).     

The effect of long term exposure to copper on physiological condition and reproduction of sheep

The influence of copper upon some physiological parameters and reproduction in ewes was studied. Four groups of animals were investigated: 1/ control ewes (untreated); 2) ewes receiving copper as a supplement over the recommended amount of copper in food (10, 25 or 50 mg Cu/ewe/day); 3/ control, superovulated ewes; and 4) ewes treated with 50 mg copper during one month and then superovulated. After 10 months of daily exposure to 10 mg of copper/ewe/day no signs of toxicity on physiological condition and reproduction were found. In ewes exposed to 25 or 50 mg of copper a decrease in blood parameters and increase in concentration of Cu in blood and liver were noticed. The wavy pattern of follicles was disturbed and disorders in fecundity, prolificacy and pregnancy occurred. Significant differences between the number of corpora lutea in superovulated control animals and experimental (Cu 50 mg) ewes were observed.     

Peripheral inhibin levels in relation to climatic variations and stage of estrous cycle in buffalo (Bubalus bubalis )

The present study investigated the peripheral plasma inhibin levels in relation to 1) the stage of estrous cycle and the effect of climatic variations. Blood samples were collected from cyclic buffalo (n=5) once daily for 32 consecutive days during the tropical hot humid (summer) and cold (winter) seasons. Estrus was recorded by parading a vasectomized bull as well as by plasma progesterone determination. In the winter season, peripheral inhibin concentrations which were lowest (0.35 +/- 0.02 ng/ml) during the mid-luteal phase of estrous cycle (Day 6 to Day 14, Day 0 = day of estrus) increased significantly (P < 0.02) to 0.47 +/- 0.04 ng/ml during the late luteal phase (Day -4 to Day -2) and then further to 0.52 +/- 0.03 ng/ml (P< 0.02) during the periestrus phase (Day -1 to Day 1). Inhibin concentrations then decreased significantly (P < 0.02) to 0.40 +/- 0.03 ng/ml during the early luteal phase (Day 2 to Day 5). In the summer season the differences in peripheral inhibin concentrations among different phases of estrous cycle were found to be nonsignificant. A comparison of the circulating inhibin concentrations between the two seasons indicated that inhibin concentrations were significantly higher in the late luteal phase (P < 0.01) and periestrus phase (P < 0.05) during the winter season compared with corresponding periods during the summer season. The present study suggests that peripheral inhibin concentrations change in the estrous cycle during cooler breeding season and that environmental heat stress can cause a reduction in peripheral inhibin concentrations.     

Impact of cold on the immune system of burying beetle,Nicrophorus vespilloides (Coleoptera: Silphidae)

Insect overwintering is one of the most astonishing phases of the insect life cycle. Despite vast amounts of knowledge available about the physiological mechanisms of this phenomenon, the impact of stress factors on insect immune system functioning during the winter is still unknown. The aim of this study is to analyze how low temperatures influence the immune system of the beetle Nicrophorus vespilloides. The results show that the beetle's immune system is differently modulated by cold induced in laboratory settings than that which occurs in natural conditions. Among beetles cultured in conditions similar to summer, low temperatures, did not influence the number of circulating haemocytes, phenoloxidase activity, haemocytes morphology, and percentage ratio of haemocyte types. In these beetles, differences were noted only in the ability of haemocytes to perform phagocytosis. Individuals acclimated in natural conditions in autumn had a higher level of humoral response and a different percentage ratio of haemocyte types. During the winter period, the number of haemocytes in the beetles decreased, but the percentage ratio of phagocytic haemocytes increased. Furthermore, we noted an increase of phenoloxidase activity. Our study also showed mitotic divisions of haemocytes in haemolymph collected from burying beetles after cold exposure and from burying beetles collected from natural conditions during autumn and winter. Differences in response to low temperatures in laboratory conditions and the natural environment suggest that the simultaneous presence of other stress factors during winter such as desiccation and starvation have a significant influence on the activity of burying beetle's immune system.     

Physiological ecology of overwintering in the hatchling painted turtle: multiple-scale variation in response to environmental stress

We integrated field and laboratory studies in an investigation of water balance, energy use, and mechanisms of cold-hardiness in hatchling painted turtles (Chrysemys picta) indigenous to west-central Nebraska (Chrysemys picta bellii) and northern Indiana (Chrysemys picta marginata) during the winters of 1999-2000 and 2000-2001. We examined 184 nests, 80 of which provided the hatchlings (n=580) and/or samples of soil used in laboratory analyses. Whereas winter 1999-2000 was relatively dry and mild, the following winter was wet and cold; serendipitously, the contrast illuminated a marked plasticity in physiological response to environmental stress. Physiological and cold-hardiness responses of turtles also varied between study locales, largely owing to differences in precipitation and edaphics and the lower prevailing and minimum nest temperatures (to -13.2 degrees C) encountered by Nebraska turtles. In Nebraska, winter mortality occurred within 12.5% (1999-2000) and 42.3% (2000-2001) of the sampled nests; no turtles died in the Indiana nests. Laboratory studies of the mechanisms of cold-hardiness used by hatchling C. picta showed that resistance to inoculative freezing and capacity for freeze tolerance increased as winter approached. However, the level of inoculation resistance strongly depended on the physical characteristics of nest soil, as well as its moisture content, which varied seasonally. Risk of inoculative freezing (and mortality) was greatest in midwinter when nest temperatures were lowest and soil moisture and activity of constituent organic ice nuclei were highest. Water balance in overwintering hatchlings was closely linked to dynamics of precipitation and soil moisture, whereas energy use and the size of the energy reserve available to hatchlings in spring depended on the winter thermal regime. Acute chilling resulted in hyperglycemia and hyperlactemia, which persisted throughout winter; this response may be cryoprotective. Some physiological characteristics and cold-hardiness attributes varied between years, between study sites, among nests at the same site, and among siblings sharing nests. Such variation may reflect adaptive phenotypic plasticity, maternal or paternal influence on an individual's response to environmental challenge, or a combination of these factors. Some evidence suggests that life-history traits, such as clutch size and body size, have been shaped by constraints imposed by the harsh winter environment.     

Ram-induced ovulation to improve artificial insemination efficiency with frozen semen in sheep.

Ram effect, defined as shortening of seasonal anestrus in ewes by exposure to the ram, is now well recognized but the underlying mechanisms are still unclear. Little information also exists whether the ram is able to influence the estrus cycle and ovulation. Three experiments were conducted to investigate endocrine response, time of ovulation and pregnancy rate of ewes in proestrus, exposed to the ram (treated) or an adult ewe (control). In the first experiment, ewes (n = 20) were treated with fluorgestone acetate pessaries for 12 days and were given eCG and cloprostenol one day before withdrawal of pessaries. On the day after removal of the pessaries ewes in the treated group (n = 10) were exposed to the ram and those in the control group (n = 10) were exposed to an adult ewe. Blood samples were taken for LH assay every 20 min from 2 h before to 24 h after ram exposure. In the second experiment, ewes (n = 120) were induced into proestrus and on the day after removal of the pessaries were exposed to either a ram (n = 60) or a ewe (n = 60) as described above and were laparoscoped 50, 60 or 70 h after pessary withdrawal (n = 20 at each time interval). In the third experiment ewes (n = 90) were induced and exposed to the ram (n = 45) or an adult ewe (n = 45) and inseminated via a laparoscope whit frozen-thawed semen at 50 or 60 h after pessary removal, respectively. Exposure to the ram was followed in 2 h by a marked rise in LH, equivalent to a preovulatory surge in duration and amplitude. It was also followed by concentrated ovulation within 25 to 30 h and by an increased pregnancy rate in exposed ewes (73.3 vs. 53.3%).     

Thermoregulation of male sheep of indigenous or exotic breeds in a tropical environment

Climate change has intensified the frequency of heat waves in the world, thereby exposing farm animals to stressful conditions. For better productive performance it is important to identify the most resilient genotypes. Thus, our objective was to evaluate the thermoregulatory responses of rams of tropical indigenous (Morada Nova and Santa Inês) and exotic breeds (Dorper and Texel), by monitoring the environmental and physiological indicators related to heat tolerance. The experiment was carried out in a tropical climate region (Cwa), in Brazil, for twelve months, which comprised spring, summer, autumn and winter. Thirty-three rams were divided into groups: Morada Nova (MN; n=8, red-coat), Santa Inês (SI; n=9, black-coat), Dorper (DO; n=8, white-coat) and Texel (TX; n=8, white-coat). The microclimatic variables were monitored, and the THI and BGHI comfort indices were calculated. Coat thickness and body surface temperatures were measured monthly, and serum triiodothyronine-T3 measurements and complete blood tests were performed. The physiological variables were evaluated every fifteen days and skin micro-biopsies were performed in the summer and winter for histological evaluation. During the warmer seasons, the THI and BGHI reached values that indicated thermal discomfort. TX showed higher coat thickness throughout the year, increased physiological variables related to thermolysis, and reduced T3 (P < 0.05). The internal temperature was permanently lower in the MN, SI and DO (P < 0.05). The body surface temperatures were affected by the coat characteristics and wool length. Hematological parameters varied in the seasons with the highest thermal conditions (P < 0.05). The MN showed larger sweat glands, while the area occupied by the sweat glands was higher in the SI. The DO showed higher hair density in the summer and winter (P < 0.05). The results indicated that the MN, SI and DO breeds overcome the thermal challenge more easily throughout the seasons due to specific adaptive morphological and physiological characteristics.