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.     

Growth Hormone Receptors in the Porcine Testis During Prepuberty

Supplementation of exogenous growth hormone (GH) during prepuberty advances onset of spermatogenesis in boars, but the mechanism of action is unknown. The present study is an investigation of the presence and characteristics of testicular growth hormone receptors (GHR). A total of 36 boars were castrated, three boars every 10 days, between the ages of 10 and 120 days. Testicular membrane preparations of 10, 20, 30, 50, 70, 100 and 120‐day‐old boars were used to determine ¹²⁵I‐bGH binding and Scatchard analysis. Liver from a 60‐kg barrow was used for comparison. Specific ¹²⁵I‐bGH binding to testicular membrane preparations occurred in all age groups with the exception of 20‐day‐old boars at levels of 30–40% of liver binding. At 30 days of age the unlabelled bGH at 1.1 ng/tube achieved half maximal inhibition (ID₅₀). Results of Scatchard analysis indicated a single class of binding sites. Binding affinity was 2.89 × 10⁹m with a binding capacity of 12 fmole/mg membrane protein. The results from this study suggest that GH may act directly on the cells of the prepubertal boar testis.     

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.     

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.     

Alteration of oceanic volcanic glass: textural evidence of microbial activity

The subsurface biosphere may constitute as much as 50 percent of Earth's biomass. Direct and indirect evidence suggests that an extensive biosphere exists in the rocks below the sea floor. This survey of basalts of the Atlantic, Pacific, and Indian Oceans supports the hypothesis that bacteria have colonized much of the upper oceanic crust, which has a volume estimated at 10(18) cubic meters. Although this is the largest habitat on Earth, its low abundance of bacteria constitutes much less than 1 percent of Earth's biomass.     

Comparison of methods used to diagnose generalized inflammatory disease in manatees (Trichechus manatus latirostris).

Manatees (Trichechus manatus latirostris) are afflicted with inflammatory and infectious disease secondary to human interaction, such as boat strike and entanglement, as well as "cold stress syndrome" and pneumonia. White-blood-cell count and fever, primary indicators of systemic inflammation in most species, are insensitive in diagnosing inflammatory disease in manatees. Acute phase-response proteins, such as haptoglobin and serum amyloid A, have proven to be sensitive measures of inflammation/infection in domestic large animal species. This study assessed diagnosis of generalized inflammatory disease by different methods including total white-blood-cell count, albumin: globulin ratio, gel electrophoresis analysis, C-reactive protein, alpha, acid glycoprotein, haptoglobin, fibrinogen, and serum amyloid A. Samples were collected from 71 apparently healthy and 27 diseased animals during diagnostic medical examination. Serum amyloid A, measured by ELISA, followed by albumin:globulin ratio, measured by plasma gel electrophoresis, were most sensitive in diagnosing inflammatory disease, with diagnostic sensitivity and specificity of approximately 90%. The reference interval for serum amyloid A is <10-50 microg/ml with an equivocal interval of 51-70 microg/ml. The reference interval for albumin:globulin ratio by plasma gel electrophoresis is 0.7-1.1. Albumin: globulin ratio, calculated using biochemical techniques, was not accurate due to overestimation of albumin by bromcresol green dye-binding methodology. Albumin:globulin ratio, measured by serum gel electrophoresis, has a low sensitivity of 15% due to the lack of fibrinogen in the sample. Haptoglobin, measured by hemoglobin titration, had a reference interval of 0.4-2.4 mg/ml, a diagnostic sensitivity of 60%, and a diagnostic specificity of 93%. The haptoglobin assay is significantly affected by hemolysis. Fibrinogen, measured by heat precipitation, has a reference interval of 100-400 mg/dl, a diagnostic sensitivity of 40%, and a diagnostic specificity of 95%.     

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.