C, N, and noble gas isotopes in grain size separates of presolar diamonds from efremovka

Nanometer-size presolar diamonds from the Efremovka CV3 chondrite were physically separated into several grain size fractions by ultracentrifugation. The coarsest size fraction is the most enriched in carbon-12; the others have broadly similar carbon isotopic compositions. Measurement of noble gases shows that their concentration decreases with decreasing grain size. This effect is attributed to ion implantation. Such an episode could occur in the envelope of a supernova that produced the diamonds, or in interstellar space; in either case, ions with energies above a certain threshold pass completely through the smaller diamond grains without being captured. Concentrations of nitrogen show only minor variations with grain size, indicating a different mechanism of incorporation into the diamonds.     

Experimental Leptospira borgpetersenii serovar hardjo infection of pregnant cattle

Objective To observe the effect upon the foetus of experimental infection of pregnant cattle with Leptospira borg-petersenii serovar hardjo .
Design A disease transmission study using pregnant cattle.
Procedure Fourteen heifers serologically negative to L hardjo were artificially inseminated and later challenged with a north-Queensland isolate of L hardjo by conjunctival inoculation. The heifers were serologically monitored and their urine examined for the presence of leptospires using culture and fluorescent-antibody tests at appropriate intervals. Elective caesarean sections were performed on pregnant heifers at 6.5 weeks after the challenge. Foetuses were examined using serological, histopathological, microbiological and fluorescent-antibody tests.
Results Ten of the heifers became pregnant, but three subsequently aborted before challenge. After challenge, all 14 heifers seroconverted and L hardjo was isolated from the urine of 6 of the 7 pregnant heifers. No evidence of foetal L hardjo infection was detected. Two of the foetuses had histopatho-logical lesions consistent with Neospora s p infection.
Conclusion It is likely that the isolate of L hardjo used in this study does not normally infect the foetus. Neospora s p may be a more significant cause of bovine reproductive wastage.     

Congenital polycystic kidney disease in lambs

AIM: To describe the pathology and inheritance of a congenital polycystic kidney disease (PKD) of sheep.

METHODS: Mode of inheritance of PKD was investigated by evaluation of results of the disorder from planned matings in two consecutive years within subsets of a flock that had a high prevalence of PKD in lambs. Gross pathological and histopath- ological studies were based on tissues derived from this study. Haematoxylin and eosin (H&E)-stained paraffin sections of kidney, liver, extrahepatic biliary and pancreatic ducts, pancreas and epididymis were used to describe the lesions.

RESULTS: Twenty-five lambs affected by PKD, of both sexes, were born, numbers in accord with those expected for an autosomal recessive disorder in the population studied. In all cases for which tissues were available, the renal, bile ductal (intrahepatic and extrahepatic), pancreatic and epididymal tissues had widespread dysplastic changes and associated cyst formation.

CONCLUSIONS: The findings of renal cysts in conjunction with cysts in other organs are unifying features in many of the human and animal forms of PKD and suggest a related pathogenic and genetic base consistent with an autosomal recessive disorder.     

Candida albicans infection in free-living populations of hihi (stitchbird; Notiomystis cincta)

Abstract

AIM: To describe the occurrence of candidiasis in hihi (stitch-bird; Notiomystis cincta) nestlings, and investigate the carriage and impact of Candida albicans infection in a free-living population of hihi.

METHODS: Mortality of nestlings was investigated in a reintroduced population of the endangered, endemic hihi at Zealandia: Karori Sanctuary, Wellington, New Zealand. Oral and faecal samples were collected from live hihi nestlings, for microbiological examination, between October 2008 and April 2009. All hihi that died and could be recovered were submitted to the New Zealand Wildlife Health Centre (NZWHC) at Massey University, for post-mortem examination. The results were compared with data obtained retrospectively from the National Wildlife Mortality (NWM) database for two other reintroduced populations of hihi on Mokoia and Tiritiri Matangi Islands.

RESULTS: Fifty chicks fledged from 82 eggs hatched during the 2008–2009 breeding season at Zealandia: Karori Sanctuary. Thirty-four live nestlings were sampled from 11 nests, and C. albicans was isolated from gastrointestinal swabs of 13 live nestlings from four nest sites. Eight (62%) of those nestlings survived to fledge, compared with 17/21 (81%) of those that tested negative (p=0.254; Fisher's exact test). Of the 32 hihi nestlings that died during the period of the study, 25 were recovered for necropsy. Histopathological examination revealed candidiasis was a factor in the deaths of four nestlings. An adult hihi that died during the period of the study at Zealandia: Karori Sanctuary was also found to have candidiasis. Retrospective analysis of data from the NWM database revealed candidiasis was also a factor in the deaths of five nestlings aged between 1 and 10 days from Mokoia Island, and of three nestlings <5 days old and one adult from Tiritiri Matangi Island.

CONCLUSIONS: Candida albicans was isolated from 38% of hihi nestlings sampled in this study, and vertical transmission of this organism from parent to offspring is likely to occur. Some colonised nestlings developed ventriculitis associated with Candida spp., but survival to fledging was not significantly different between nestlings that tested positive or negative, although the fate of birds following fledging was unknown.     

Effects of Egg Yolk and Cooling Rate on the Survival of Refrigerated Red Deer (Cervus elaphus hispanicus) Epididymal Spermatozoa

Egg yolk is a common component to sperm refrigeration for most of the deer species, the role of which is to protect sperm membranes against cold shock. In addition, there have been many studies of conservation of ejaculated semen from stags, but few have been reported for epididymal spermatozoa. This work was designed to investigate the combined effects of cooling rates (slow: 0.23 degrees C/min vs rapid: 4.2 degrees C/min) from room temperature to 5 degrees C, and egg-yolk concentration (0, 5 or 20%) in the extender on the survival of Iberian red deer epididymal spermatozoa refrigerated at 5 degrees C. Heterospermic sperm samples were diluted to a final sperm concentration approximately 400x10(6) sperm/ml with a Tris-citrate-fructose (TCF)-egg-yolk diluent. Sperm quality was in vitro judged by microscopic assessments of individual sperm motility [sperm motility index (SMI)], and of plasma membrane (hypo-osmotic swelling test) and acrosome (NAR) integrities. Our results first showed that the presence of egg yolk in the extender significantly improves (p=0.01) the viability and sperm motility after sperm dilution. In addition, acrosome and plasma membrane integrities post-refrigeration did not differ significantly between cooling procedures; however, the SMI differed significantly between cooling procedures (slow: 46.6% vs rapid: 50.0%; p=0.01). Our results also showed that sperm quality was significantly (p<0.01) affected by the combined effects of egg-yolk concentration and cooling procedure, being rapid cooling with 20% of egg yolk the most suitable combination for epididymal sperm refrigeration. In conclusion, egg-yolk improved red deer epididymal spermatozoa characteristics after dilution. Rapid cooling protocol using TCF with 20% egg-yolk significantly improved sperm motility of red deer epididymal spermatozoa after cooling.     

Does the Amount of Progesterone in Intravaginal Implants Used to Synchronise Oestrus Affect the Reproductive Performance of Brahman Heifers Artificially Inseminated at a Fixed Time

The study tested the hypothesis that reduced intravaginal implant progesterone (P₄) concentration to synchronise oestrus would increase pregnancy rates to fixed‐time artificial insemination (FTAI) in Bos indicus heifers. Brahman heifers (n = 294; 2 year) were body condition scored (BCS), weighed and scanned for presence of a corpus luteum (CL). Only cyclic heifers were selected and allocated randomly within BCS and 25 kg bodyweight category to one of three P₄ treatment groups. On day 10, heifers received a P₄ implant (CueMate‐1‐pod, 0.78g P₄; CueMate‐2‐pod, 1.56g P₄; or CIDR‐B, 1.9g P₄), 2 mg oestradiol benzoate (ODB) intramuscularly (IM) and 250 ug cloprostenol IM. At day 2, the implant was removed, 250 ug cloprostenol was injected IM and tail paint applied. The heifers received 1 mg ODB 24 h later and were FTAI 48–54 h after implant removal (day 0). Ten randomly selected heifers per group were blood sampled and scanned at days 10, 2, 0 and 6 to define the P₄ profiles pre‐ and post‐FTAI. Heifers were heat‐detected 18–20 days post‐FTAI and oestrous heifers AI’d by the AM/PM rule. Bulls joined the heifers on day 27 post‐FTAI. Transrectal ultrasonography estimated conception date on day 72. Statistical analysis examined the effects of treatment, technician, semen, ovarian status, BCS and liveweight, on pregnancy rate (PR) to FTAI. There was no significant difference (p = 0.362) in PR between treatment groups (CueMate 1‐pod, 36.4%; CueMate 2‐pod, 39.6%: CIDR‐B, 28.3%), but PR was higher in those heifers with increased BCS between FTAI and pregnancy diagnosis (p = 0.005). Thirty‐three per cent of monitor heifers had plasma P₄ concentrations of <1 ng/ml on day 6 after FTAI; only 20% of these conceived vs 60% of heifers with P₄ ≥ 1 ng/ml. In summary, no significant difference in PR was identified between treatments but good BCS and a rising plane of nutrition were critical to PR of these pure grade Brahman heifers in northern Australia.     

Identification of Sperm-Head Morphometric Subpopulations in Iberian Red Deer Epididymal Sperm Samples

Computer-assisted sperm morphometry analysis (CASMA) was used in this study to identify sperm morphometric subpopulations in Iberian red deer epididymal sperm samples. Epididymal sperm samples were collected from 37 mature stags and were divided. One portion was diluted in a Tris–citrate–egg yolk medium. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours using a conventional protocol. After thawing, sperm smears were prepared as described for extended samples. All slides were air-dried and stained with Hemacolor^®. The sperm-head dimensions for a minimum of 145 sperm-heads were analyzed from each sample by means of the Sperm-Class Analyser^®, and the mean measurements recorded. Each sperm-head was measured for four primary sperm-head parameters, and five parameters of head shape. All sperm morphometric parameters evaluated were placed in a statistical database and a multivariate cluster analysis was performed. The clustering analyses, based on 10 867 individual spermatozoa, revealed the existence of three subpopulations (SP₁, SP₂, SP₃) of spermatozoa with different morphometric characteristics (p  <  0.001). The proportion of spermatozoa present in any of the three subpopulations remained constant (p  >  0.05) through the cryopreservation process. Pre-freeze and post-thaw sperm quality was in vitro evaluated by microscopic assessments of individual sperm motility and of plasma membrane and acrosome integrities. In conclusion, our results show that applying the CASMA techniques and multivariate cluster analyses, it was possible to determine that three subtle subpopulations of spermatozoa with different morphometric characteristics coexist in red deer semen.     

Development of a Multiplex PCR for the Identification of Major Pathogenic Bacteria of Post‐Partum Endometritis in Dairy Cows

Post‐partum period has an important role in cows' breeding due to its effects on reproductive efficiency and subsequent pregnancy. Escherichia coli, Trueperella pyogenes (Arcanobacterium pyogenes), Fusobacterium necrophorum and Prevotella melaninogenicus are recognized as major pathogens associated with uterine endometrial lesions. The objective of this study was to identify these pathogens using the polymerase chain reaction (PCR) as a culture‐independent sensitive method. A total of 172 cows were examined 25–35 days post‐partum, and 128 cows were examined at 2 weeks later (39–49 days post‐partum). Uterine discharges were collected by covered plastic infusion pipettes. The prevalence of endometritis was greater in the first examination than the second (35.5% vs. 16%). E. coli was detected in eight of the samples, T. pyogenes was detected in 13 of the samples and F. necrophorum was detected in 11 of the samples. There was no positive sample of P. melaninogenicus. Uterine contamination by T. pyogenes and F. necrophorum in the first examination was higher than the second examination. T. pyogenes affected as a tendency the prevalence of clinical endometritis in first examination. Primiparous cows showed 4.02 times higher odds of clinical endometritis compared with second‐parity cows in first examination. A multiplex PCR protocol as a simple, less expensive, fast assay was introduced to identify E. coli, T. pyogenes and F. necrophorum.