Identification of Corynebacterium pseudotuberculosis from sheep by PCR-restriction analysis using the RNA polymerase β-subunit gene (rpoB)

Caseous lymphadenitis, caused by Corynebacterium pseudotuberculosis, has a high prevalence in many regions of the world, including Argentina and Brazil. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for the identification of this microorganism was designed based on the hypervariable region of the polymorphic RNA polymerase β-subunit gene (rpoB). All available CorynebacteriumrpoB sequences were analyzed by computer-assisted restriction analysis. The rpoB PCR-RFLP pattern predicted by using endonucleases MseI and StuI clearly differentiated C. pseudotuberculosis from sixty-one other Corynebacterium species. This method was successfully applied to identify twelve wild C. pseudotuberculosis ovine isolates and one caprine isolate. It was also used to differentiate C. pseudotuberculosis from Arcanobacterium pyogenes, an ovine pathogen with similar clinical characteristics. These results indicate that this new molecular method can be used for the reliable identification of the pathogen, essential for the timely detection of infected animals and for epidemiological studies.     

Cloning and phylogenetic analysis of Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) from Indian dromedaries (Camelus dromedarius)

The cDNAs of two proinflammatory cytokines viz., IL-6 and TNF-α from dromedarian camels were amplified by PCR using bactrian camel sequences and subsequently cloned for sequence analysis. Relationship based on amino acid revealed that dromedarian camel IL-6 shared 99.5% identity both at nucleotide and amino acid level with bactrian camel IL-6 and in case of TNF-α, the identity of dromedarian camel was 99.4% and 99.1% at nucleotide and amino acid level, respectively with that of bactrian camel. Phylogenetic analysis based on their amino acid sequences indicated the close relationship in these cytokine genes between dromedarian camel and other members of camelids.     

Cervico-vaginal mucus (CVM) – an accessible source of immunologically informative biomolecules

Cervico-vaginal mucus (CVM), the product of epithelial cells lining the uterus, cervix and vagina, is secreted to facilitate uterine lubrication and microbial clearance. Predominantly composed of water and mucins, CVM also contains high levels of immuno-active proteins such as immunoglobulin A (IgA), lactoferrin and lysozyme which protect against infection by blocking adhesion and mediating microbial killing. The repertoire of cytokines, chemokines and antimicrobial peptides is predominantly generated by the secretions of endometrial epithelial cells into the uterine lumen and concentrated in the CVM. The quantity and relative proportions of these inflammatory biomarkers are affected by diverse factors including the estrus cycle and health status of the animal and therefore potentially provide important diagnostic and prognostic indicators. We propose that measuring molecular signatures in bovine CVM could be a useful approach to identifying and monitoring genital tract pathologies in beef and dairy cows.     

Turritella declivis Adams & Reeve,in Reeve, 1849 (Mollusca: Gastropoda) – a South African not an Australian species,and a characteristic component of the Agulhas Bank benthos

The name Turritella declivis, as described by Adams and Reeve in the ‘The Zoology of the Voyage of HMS Samarang’ (Adams & Reeve 1848-1850), is currently used for two distinct species of turritellid gastropod, one inhabiting the Agulhas Bank off South Africa and the other occurring off the Australian coast. Examination of the type material clearly shows that the name is referable to the southern African species. The Australian species must henceforth be known as Turritella captiva Hedley, 1987, a name previously regarded as a junior synonym of T. declivis. The original type locality ‘China Seas’ is erroneous, as was often the case with material collected during the voyage of the Samarang. It is emended to be the Agulhas Bank, over much of which T. declivis is common and may, in places, be hugely abundant, forming Turritella-dominated benthic communities. Such communities have been recorded in other parts of the world and are frequently associated with seasonal upwelling events, but have not previously been documented off South Africa.     

Extraction of 22 equine cheek teeth with displaced sagittal fractures using polymethylmethacrylate stabilisation (2011–2016)

Displaced sagittal cheek tooth fractures are a cause of oral pain, quidding and apical infection. Intraoral extraction is the preferred technique to remove affected teeth, but can be difficult due to displaced and friable fracture fragments. Stabilising fracture fragments via filling of the fracture space with polymethymethacrylate (PMMA) prior to removal may be a useful method to facilitate intraoral extraction. Case details were examined retrospectively. A total of 22 cheek teeth required extraction in 20 horses because of displaced sagittal fractures. Clinical diagnoses were made using oral examination, oral endoscopy, skull radiography and computed tomography. All procedures were performed in standing, sedated horses in stocks. Fracture spaces were cleaned and packed with PMMA and teeth removed using a routine intraoral extraction technique. Digital photographs of extracted teeth were taken and tooth measurements calibrated using digital image software. Intraoperative difficulties, as well as post-operative complications were recorded. A total of 21 maxillary and one mandibular cheek teeth were extracted. All maxillary teeth had advanced infundibular caries. Intraoral extraction was successful in 16 cases; six were unsuccessful and required repulsion due to tooth fragmentation or abnormal dental anatomy. In 11 cases, maxillary or conchofrontal sinus trephination was performed to either treat sinusitis, repulse the tooth, or both. Two horses developed short-term complications following local anaesthesia of the maxillary nerve. The mean ratio of fracture depth to tooth length was 0.59 and mean ratio of fracture width to tooth width 0.53. The limitations of the study are its small sample size, retrospective nature and lack of control group to compare extraction success in PMMA and non-PMMA groups. It was concluded that using PMMA to stabilise displaced sagittal fractures in equine cheek teeth is a simple, effective method of facilitating intraoral extraction and may reduce the need for more invasive procedures.     

Nonsurgical removal of urethral uroliths using a self-retaining retractor with elastic stays in female guinea pigs (Cavia porcellus): 16 Cases (2006–2019)

Sixteen cases of client-owned female guinea pigs (Cavia porcellus) involved urethrolith removal directly from the urinary orifice or urethra with the aid of a lightweight self-retaining retractor ring with elastic stays. The procedure was performed in fifteen guinea pigs and repeated once due to reoccurrence of urolith formation. Urethrolithiasis was diagnosed by direct palpation of the urethrolith, and in all but one case confirmed radiographically. Uroliths were removed whole or in sections after direct visualization with the help of the retractor. Only minor complications associated with the procedure were observed and most patients survived to discharge. This is the first case series to describe the use of a self-retaining retractor to facilitate simple nonsurgical removal of urethroliths in female guinea pigs. This procedure is rapid, minimally invasive, technically simple, and cost effective in female guinea pigs with uroliths in the urinary orifice or distal urethra.     

Canola meal as an alternative dietary protein source in quail (Coturnix coturnix) diets – A review

ABSTRACT

Fast-growing and highly adaptable avian species such as the Japanese quail (Coturnix coturnix japonica) have greater potential for use in alleviating food and nutrition insecurity. However, the continued use of soybeans as a protein source may reduce the contribution of quails to food and nutrition security. It is, therefore, prudent that alternative dietary proteins, with no direct food value for humans, be evaluated to allow for sustainable intensification of quails. One such protein source is canola meal, a by-product of oil extraction from canola seed. This review explores the potential of canola meal as a replacement for soybean meal in quail diets by presenting a comparative analysis of the composition and functional properties of the two proteins. A balanced assessment of in vivo responses to canola meal in quails and other avians is presented with the objective of enhancing the quail's contribution to food and nutrition security.     

Molecular structure, phylogenetic analysis, tissue distribution, and function characterization of interferon-γ-inducible lysosomal thiol reductase (GILT) gene in sheep (Ovis aries)

Interferon-γ-inducible-lysosomal thiol reductase (GILT) plays a key role in the processing and presentation of MHC class II-restricted antigen (Ag) by catalyzing disulfide bond reduction. In this study, a sheep cDNA, sGILT, encoding GILT protein was isolated from the spleen cDNA library of Ovis aries. It codes for a deduced protein of 244 amino acids with a putative molecular weight of 27.6 kDa, which has all the typical structural features of GILT protein including an active-site CXXC motif, a GILT signature sequence CQHGX(2)ECX(2)NX(4)C, and 5 conserved cysteines. Sequence comparison indicated the amino acid sequence of sGILT showed high identity to cow GILT (93.03%). Phylogenetic analysis showed that sGILT and cow GILT shared the greatest homology. The result of real-time PCR suggested that sGILT mRNA was expressed in a tissue-specific manner and obviously up-regulated in splenocytes and PBMCs after induction with lipopolysaccharide (LPS). Recombinant sGILT fused with His(6) tag was efficiently expressed in Escherichia coli BL21 (DE3) and purified by Ni-NTA affinity chromatography. Its expression was confirmed by SDS-PAGE and Western blotting analysis. Thiol reductase activity was assessed using antibody as the substrate. These results suggest that sGILT has the activity of disulfide bond reduction and indicate that sheep also express a protein that has been found to maintain first line of innate immune defense at basal level.     

Detection of Subclinical Mastitis in Dromedary Camels (Camelus dromedarius) using Somatic Cell Counts and the N-Acetyl-β-D-glucosaminidase Test

Somatic cell counts, N-acetyl--D-glucosaminidase (NAGase) activity and the infection status of the udder were determined in quarter milk samples (n = 86) from 22 multiparous, clinically healthy camels, traditionally managed by Bedouin nomads in the Negev desert, Israel. Seventy (81.4%) of the 86 samples examined contained bacteria, of which 35 (40.7%) gave mixed isolations of two or more bacteria, suggesting the existence of subclinical mastitis in the camel herds studied. Sixteen samples (18.6%) yielded no growth of bacteria. Staphylococcus aureus, Micrococcus spp., Bacillus spp., Streptococcus dysgalactiae and Escherichia coli were the main organisms isolated. The somatic cell count (SCC) ranged from 1.01×10⁵ to 11.78×10⁶ cells/ml. NAGase values were between 41.4 and 372 NAGase units. Quarter milk samples that contained bacteria had significantly (p<0.01) higher mean values for SCC but the mean NAGase levels were not significantly different for the bacteriologically negative and positive samples. There was a low correlation coefficient (r ² = 0.097) between the SCC and NAGase in the quarter milk samples from which bacteria were not isolated (n = 16) and a low negative correlation (r ² = –0.038) with the samples that contained bacteria (n = 70). The type of bacteria had a significant effect (p<0.01) on the SCC but not on the NAGase activity. Quarter samples from which Staphylococcus aureus (coagulase positive) was isolated showed the highest mean SCC and this organism is therefore suspected to be the underlying cause of the subclinical mastitis. The SCC gave a better indication of the presence of pathogenic microorganisms in milk samples than did NAGase.     

Determination of flagellar types by PCR-RFLP analysis of enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing E. coli (STEC) strains isolated from animals in São Paulo, Brazil

This study evaluated the polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) analysis of fliC for typing flagella antigen (H) of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains isolated from different animals. The molecular typing of the H type was efficient in the determination of 93 (85%) strains. Two nonmotile (H-) E. coli strains showed a PCR-RFLP electrophoretic profile that did not match known H type patterns. The fliC nucleotide sequence of strains B2N and 4a revealed a nucleotide substitution at the restriction site and a nucleotide insertion that generated a stop codon, respectively. The results of this study showed that PCR-RFLP analysis of fliC is faster, less laborious and as efficient for the determination of H type E. coli isolated from animals, compared to serotyping and that it is useful in determining H type in nonmotile strains and strains expressing non-reactive H antigens. Moreover, the fliC sequence of strain B2N suggests that we could have found a new flagellin antigen type.     

Transcriptome analysis of the effect of pyrroloquinoline quinone disodium(PQQ·Na_2) on reproductive performance in sows during gestation and lactation

Background: Pyrroloquinoline quinone(PQQ), which is a water soluble, thermo-stable triglyceride-quinone, is widely distributed in nature and characterized as a mammalian vitamin-like redox cofactor. The objective of this study was to investigate the effects of pyrroloquinoline quinone disodium(PQQ·Na2) on reproductive performance in sows.Results: Dietary supplementation with PQQ·Na2 significantly increased the total number of piglets born, the number of piglets born alive and the born alive litter weight. It also increased the antioxidant status in the placenta, plasma and milk. The concentration of NO was significantly increased in the plasma and placenta. RNA-seq analysis showed that462 unigenes were differentially expressed between the control(Con) treatment and PQQ treatment groups.Among these unigenes, 199 were upregulated, while 263 unigenes were downregulated. The assigned functions of the unigenes covered a broad range of GO categories. Reproduction(27, 7.03%) and the reproduction process(27, 7.03%) were assigned to the biological process category. By matching DEGs to the KEGG database, we identified 29 pathways.Conclusions: In conclusion, dietary supplementation with PQQ·Na2 in gestating and lactating sows had positive effects on their reproductive performance.