Contrasting effects of heat shock during in vitro maturation on development of in vitro‐fertilized and parthenogenetic bovine embryos

This study investigated the influence of heat shock during in vitro maturation on embryo development following in vitro fertilization (IVF) or parthenogenesis (Part). Immature bovine cumulus–oocyte complexes were exposed to heat shock (41.0°C) during the first 12 hr of in vitro maturation (IVM), followed by 12 hr at 38.5°C. Control group consisted of in vitro maturation for 24 hr at 38.5°C. Oocytes were in vitro‐fertilized or activated with ionomycin and cultured in vitro for 192 hr post‐in vitro insemination or parthenogenetic activation (hpia). There was an interaction (p < .01) between temperature of IVM and method of oocyte activation (IVF or Part) for cleavage at 48 hpia. Heat shock had a negative impact (p < .01) on cleavage of IVF embryos, whereas no (p > .05) effect was found in the Part embryos. Embryo development towards blastocyst stage at 168 and 192 hpia decreased in both IVF and Part embryos derived from heat‐shocked oocytes. Heat shock increased (p < .05) the apoptotic index in Part blastocysts, but no effect (p > .05) was found in IVF counterparts. Heat shock also down‐regulated the expression of AQP3 (p < .01) and up‐regulated the expression of HSP70.1 (p < .01) in Part blastocysts, whereas it down‐regulated the expression of ATP1A1 (p < .05) in IVF blastocysts. In conclusion, the effects of heat shock during IVM on early embryo cleavage and blastocyst apoptosis are influenced by the method of oocyte activation and expression of some genes can be disturbed in embryos derived from heat‐shocked oocytes.     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Effects of intensification of pastoral farming on greenhouse gas emissions in New Zealand

Abstract

AIMS: The aims of this cross-sectional study were to investigate the herd and cow-level prevalence of bovine digital dermatitis (BDD) in dairy farms in the northern Taranaki region of New Zealand, and to identify whether there was any spatial clustering of herds with the disease.

METHODS: A survey of 224 dairy farms in the northern Taranaki region of New Zealand was undertaken from September 2014 to February 2015. Following training in robust criteria to confirm BDD visually, a technician inspected the rear feet of every milking cow on the farms during milking. The identity of cows with lesions and the feet involved were recorded. The proportion of cows affected among the inspected population (cow-level prevalence), the proportion of a herd affected (farm-level prevalence), and proportion of farms with ≥1 cow with lesions, were calculated. A bivariate K function analysis was then used to assess whether farms with ≥1 cow with lesions were clustered, after accounting for the distribution of the farms involved in the study.

RESULTS: Bovine digital dermatitis lesions were observed on 143/224 (63.8 (95% CI=57.5–70.1)%) farms. Within-farm prevalence was 0% on 81 (36.2%) farms, between >0 and <3% on 120 (53.5%) farms, with a maximum prevalence of 12.7% on one farm. Overall, cow-level prevalence was 707/60,455 (1.2 (95% CI=0.9–3.0)%), and on affected farms was 707/41,116 (1.7 (95% CI=1.4–2.1)%). In affected cows, 268/707 (37.9%) had a lesion on left foot only, 262/707 (37.1%) on the right foot only and 177/707 (25.0%) on both feet. The K function analysis showed no evidence of clustering of farms with BDD.

CONCLUSIONS: Bovine digital dermatitis was widespread among the survey farms, but there was no evidence that there was any clustering of herds with BDD. The cow-level prevalence on affected farms was much lower than reported elsewhere.

CLINICAL RELEVANCE: Although the prevalence at the cow level was low, if these data are representative of other regions of New Zealand, BDD could easily become a major problem on dairy farms in New Zealand, as has been observed in other countries.     

Fertility of Holstein cows and heifers submitted to timed artificial insemination and receiving one or two doses (12 h apart) of semen

The objective of this retrospective study was to assess the effect of receiving a single (n = 50,285) or double (n = 4392) artificial insemination (AI), 12 h apart, within a timed artificial insemination protocol on pregnancy per AI (P/AI) in nulliparous heifers (inseminated with either sex-sorted or conventional semen) and pluriparous Holstein cows in a commercial dairy herd. Also, this study aimed to investigate the relationship between temperature-humidity index (THI) and time of the first AI and fertility. Fertility of cows receiving two AI with normothermia (THI <68) was higher (p < .05) than cows receiving a single AI (42.9% vs. 36.4%). P/AI of cows receiving two AI with severe heat stress (THI >85) was higher (p < .05) than cows receiving a single AI (21.0% vs. 12.6%). Regardless of heat stress conditions, applying the first AI in the morning increased (p < .05) P/AI in cows with double AI than in cows whose first AI occurred in the afternoon (38.4 vs. 33.3%). With moderate heat stress, and sexed-sorted semen, P/AI to timed AI was higher (65.0 vs. 51.9%; p < .05) in heifers receiving double AI than those serviced once. It was concluded that double AI, 12 h apart, enhanced fertility at timed AI than herd mates with a single AI, particularly with heat stress at breeding.     

Ovicidal activity of seven <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> fungal isolates on <Emphasis Type="Italic">Ascaris suum</Emphasis> eggs

The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water–agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p > 0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.     

Virulence factors of avian pathogenic Escherichia coli isolated from broilers from the south of Brazil

Sixty-three Escherichia coli strains isolated from broilers with respiratory problems were examined for virulence factors, hemolysin synthesis ability, motility, hemagglutination capacity, operon pap presence, colicin production, and serum resistance. The capacity to hemagglutinate guinea pig erythrocytes was found in 53 (84.1%) of the samples, but only 30 (47.6%) agglutinated chicken erythrocytes. D-mannose-sensitive hemagglutination against guinea pig erythrocytes was found in 19 (30.2%) samples and against chicken erythrocytes, in 15 (23.8%) samples, whereas the D-mannose-resistant hemagglutination with guinea pig erythrocytes was found in 34 (54%) samples, and 13 of these (20.6%) showed this characteristic against chicken erythrocytes. Operon pap, P fimbria codifier, was detected in 26 samples in a total of 34 D-mannose-resistant samples. Colicin production was observed in 55 (87.3%) of the strains, and 41.8% presented V colicin production. Of the samples analyzed, 56 (88.9%) presented serum resistance, six (9.5%) were intermediate, and only one (1.6%) was sensitive to the action of the complement. The diversity of virulence profiles detected in the samples in this study explains in part the multifactorial characteristics of avian colibacillosis.     

First Isolation of Rotavirus Associated with Neonatal Diarrhoea in Guanacos (Lama guanicoe) in the Argentinean Patagonia Region

Group A rotavirus (RV) and coronavirus (CV) are common viral pathogens associated with neonatal diarrhoea in numerous animal species. The purpose of this work was to investigate the presence of these viral agents in two farm populations of captured guanacos (Lama guanicoe) in the Argentinean Patagonia region, that developed severe diarrhoea outbreaks. Stool and serum samples were analysed for RV and bovine CV antigen and antibody by enzyme‐linked immunosorbent assay. Rotavirus was detected in faeces from two new‐born guanacos with acute diarrhoea, one in each farm. After electrophoretic analysis, each isolated strain, showed a distinctive long dsRNA electropherotype characteristic of group A rotaviruses (4:2:3:2). In addition, 95% (38 of 40) of the sampled animals were positive for RV antibodies, suggesting a high prevalence of RV infection in the populations tested. No evidence of CV circulation by antigen or antibody analysis was observed. To our knowledge, this is the first report of the detection and isolation of RV associated with neonatal diarrhoea in Lama guanicoe.     

Serum biochemical analytes in captive Amazonian manatees (Trichechus inunguis)

Background: Establishment of reference values for serum biochemical analytes is important for monitoring health and physiological status of captive animals. Objective: The purpose of this study was to measure and report ranges for serum biochemical analytes in Amazonian manatees (Trichechus inunguis). Methods: Blood samples were collected from 24 healthy captive Amazonian manatees that comprised a mixture of adults, subadults, and calves and males and females; serum analytes were measured and analyzed using a dry reagent bench‐top chemical analyzer. Comparisons were made between sexes and with previously published values of closely related species. Results: Medians and ranges (minimum–maximum) of values for the analytes were: lactate dehydrogenase (LDH), 151 (111–278) U/L (n=20); creatine kinase, 144 (76–315) U/L (n=11); alanine aminotransferase, 10 (2–28) U/L (n=18); aspartate aminotransferase, 14 (5–28) U/L (n=21); γ‐glutamyltransferase, 47 (36–73) U/L (n=21); amylase, 1428 (1010–1874) U/L (n=21); alkaline phosphatase, 73 (36–141) U/L (n=19); total protein, 6.8 (6.2–8.0) g/dL (n=24); albumin, 3.3 (2.6–4.1) g/dL (n=21); cholesterol, 188 (101–399) mg/dL (n=21); triglycerides, 126 (60–236) mg/dL (n=21); glucose, 47 (22–69) mg/dL (n=21); urea, 43 (21–69) mg/dL (n=21); uric acid, 1.1 (0.5–1.8) mg/dL (n=22); creatinine, 2.2 (1.5–3.3) mg/dL (n=22); total bilirubin, 0.2 (0.2–2.0) mg/dL (n=21); calcium, 12.7 (10.2–18.6) mg/dL (n=24); iron, 282 (207–457) μg/dL (n=13); and magnesium, 6.9 (4.3–8.9) mg/dL (n=20). With the exception of LDH, no differences were observed between sexes. Conclusions: The ranges obtained in this study provide important preliminary estimates for concentrations and activities of serum analytes in Amazonian manatees until a larger reference interval study can be conducted.