Molecular characterisation of Cryptosporidium isolates from pet reptiles

In this study, we investigated the presence of Cryptosporidium in 171 faecal samples from reptiles commonly used as pet animals. These include lizards belonging to the genera Eublepharis, Pogona, Chlamydosaurus, Hemiteconyx, Teratoscincus, Tiliqua, Iguana, and Chamaeleo, snakes of the genera Lampropeltis, Elaphe, Python, Boa and Corallus, and tortoises belonging to the genera Testudo and Kinixys. Cryptosporidium oocysts were detected by immunofluorescence using a commercially available kit and cryptosporidial DNA by amplification of a polymorphic fragment of the 18S rDNA and the HSP70 locus.Cryptosporidium was detected in 38.6% and 25.1% of the samples analysed by immunofluorescence and PCR, respectively. Molecular characterisation of the isolates confirmed that C. serpentis and C. varanii (syn. C. saurophilum) are the main species involved in infection in pet reptiles but also showed the presence of C. parvum and C. muris, as well as other species or genotypes of this parasite including the Cryptosporidium mouse genotype and Cryptosporidium tortoise genotype previously described in reptiles. In addition, a Cryptosporidium sp. was isolated from a chameleon and a python.     

Identification and characterization of lactic acid bacteria isolated from mixed pasture of timothy and orchardgrass,and its badly preserved silage

In order to understand the relationship between lactic acid bacteria (LAB) species and silage fermentation, a total of 65 LAB strains isolated from mixed pasture of timothy (Phleum pratense L.) and orchardgrass (Dactylis glomerata L.), and its badly preserved silages were subjected to phenotypic and genetic analysis. According to these analyses, the isolates were divided into 13 groups, including Enterococcus gallinarum, Lactobacillus acidipiscis, L. coryniformis subsp. coryniformis, L. coryniformis subsp. torquens, L. curvatus, L. paraplantarum, L. plantarum subsp. argentoratensis, L. plantarum subsp. plantarum, L. sakei subsp. carnosus, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Leuconostoc pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Weissella hellenica, Weissella paramesenteroides and Carnobacterium divergens. This is the first report to document that C. divergens, L. acidipiscis, L. sakei subsp. carnosus, L. garvieae, phenotypically novel L. lactis subsp. cremoris, E. gallinarum and W. hellenica are present in vegetative forage crops. L. plantarum group strains were most frequently isolated from the badly preserved silages. Some isolates showed a wide range of growth preferences for carbohydrate utilization, optimal growth pH and temperature in vitro, indicating that they have a high growth potential. These results are useful in understanding the diversity of LAB associated with decayed silage of timothy and orchardgrass.     

High‐level exogenous trans10, cis12 conjugated linoleic acid plays an anti‐lipogenesis role in bovine mammary epithelial cells

Primary bovine mammary epithelial cells (BMECs) were treated by 0, 37.5, 75, 112.5, 150 μmol/L trans10, cis12 conjugated linoleic acid (CLA) to evaluate the effects of different level trans10, cis12 CLA on lipogenesis in BMEC. Addition of 75–150 μmol/L trans10, cis12 CLA reduced significantly the triacylglycerol (TAG) content (P < 0.05), but did not have inhibiting action on cell proliferation (P > 0.05). Treatment with 150 μmol/L trans10, cis12 CLA for 48 h resulted in a 17.1% reduction (P < 0.0001) of medium chain fatty acids (MCFA, C14 < C < C16), a 26.5% reduction (P < 0.0001) of unsaturated fatty acids (UFA) and a corresponding reduction of the mRNA abundance of acetyl coenzyme A (acetylCoA) carboxylase (ACC) (P = 0.046), fatty acid synthase (FAS) (P = 0.017) and stearoylCoA desaturase1 (SCD1) (P = 0.002). Another finding was that trans10, cis12 CLA elevated expression of diacylglycerol acyltransferase2 (DGAT2) (P = 0.020) and long chain acylCoA synthetases (ACSL) (P = 0.032). In conclusion, higher trans10, cis12 CLA, not low trans10, cis12 CLA, inhibited milk fat synthesis and changed fatty acid composition by regulating the expression of FAS, ACC, SCD1, DGAT2 and ACSL.     

PCR‐based Detection of Genes encoding Virulence Determinants in Staphylococcus aureus from Birds

The present study was designed to comparatively investigate 19 Staphylococcus aureus strains isolated from specimens of 19 different birds during routine microbiological diagnostics. The S. aureus strains were characterized genotypically by polymerase chain reaction (PCR) amplification using 62 different oligonucleotide primers amplifying genes encoding staphylococcal cell surface proteins, exoproteins and two classes of the accessory gene regulator agr. All 19 investigated S. aureus were positive for the gene segment encoding a S. aureus‐specific part of the 23S rRNA, the genes encoding thermostable nuclease (nuc), clumping factor (clfA) and coagulase (coa) and the gene segments encoding the Xr‐repetitive region and the immunoglobulin G (IgG)‐binding region of protein A (spa). In addition, all tested strains were positive for the genes hla and fnbA and negative for the genes seb, sec, sed, see, sej, tst, eta and etb. The remaining genes, including sbi, hlb, fnbB, ebpS, cna (domains A and B), cap5, cap8, set1, agr class I, agr class II, sea, seg, seh and sei were detected in a variable number of isolates. The presented data give an overview on the distribution of virulence determinants of S. aureus strains isolated from birds. This might be useful to understand the role of these virulence determinants in bird infections.     

Basic characterization of avian β‐defensin genes in the Japanese quail,Coturnix japonica

In this study, we identified a cluster of 14 avian β‐defensins (AvBD; approximately 66 kbp) in the Japanese quail, Coturnix japonica. Except for AvBD12 (CjAvBD12) and ‐13, the CjAvBDs coding sequences exhibited greater than 78.0% similarity to the respective orthologous chicken AvBD genes (GgAvBD). The putative amino acid sequence encoded by each CjAvBD contained six cysteine residues and the GXC (X1‐2) motif considered essential for the β‐defensin family. Each CjAvBDs also formed a sub‐group with the respective orthologous genes of various bird species in a phylogenetic tree analysis. Synteny between the CjAvBD cluster and GgAvBD cluster was confirmed. The CjAvBD cluster was mapped on the long‐arm end of chromosome 3 by linkage analysis based on single nucleotide polymorphisms (SNPs) of CjAvBD1 and CjAvBD12 (approximately 46kbp), as well as GgAvBD cluster. We also confirmed that CjAvBD1, ‐4, ‐5, ‐9, and ‐10 are transcribed in 20 tissues, including immune and digestive tissues. However, our experimental data indicated that the CjAvBD cluster lacks the AvBD3 and ‐7 loci, whereas the CjAvBD101α, ‐101β, and ‐101θ loci arose from gene duplication of the AvBD6 orthologous locus in the CjAvBD cluster after differentiation between Coturnix ‐ Gallus.     

Buffer nitrogen solubility,in vitro ruminal partitioning of nitrogen and in vitro ruminal biological activity of tannins in leaves of four fodder tree species

This study explores the chemical composition, buffer N solubility, in vitro ruminal N degradability and in vitro ruminal biological activity of tannins in leaves from Gliricidia sepium, Leucaena leucocephala, Morus alba and Trichanthera gigantea trees. These tree leaves are a potential protein source for ruminants, but their site‐influenced nutritive value is largely unknown. Leucaena leucocephala leaves had the highest N content (42.1 g/kg DM), while T. gigantea leaves had the least (26.1 g/kg DM). Leucaena leucocephala had the highest buffer solubility index (20%), while 10% of the total N in leaves of the other three species was soluble. The rapidly fermentable N fraction ‘a’ was highest in M. alba leaves (734.9 g/kg DM) and least in T. gigantea leaves (139.5 g/kg DM). The rate of fermentation (c) was highest for M. alba (7%/hours) leaves. No significant correlations were recorded between buffer solubility index of N and in vitro ruminal N degradability parameters: a, b, and c. The highest response to tannin inactivation using polyethylene glycol, in terms of percentage increase in 36‐hours cumulative gas production, was recorded in M. alba (39%) and T. gigantea (38%) leaves. It was concluded that buffer solubility of N is not a good indicator of ruminal N degradation in the leaves of these tree species. Leaves of M. alba could be more valuable as a source of rapidly fermentable N when animals are offered low‐protein, high‐fibre diets compared with other tree species evaluated in the current study. However, when feeding M. alba leaves, the role of tannins must be considered because these secondary plant compounds showed significant in vitro ruminal biological activity.     

Synergistic and antagonistic hemolytic activities of bacteria of genus Arcanobacterium and CAMP-like hemolysis of Arcanobacterium phocae and Arcanobacterium haemolyticum with Psychrobacter phenylpyruvicus

A total of 57 bacteria representing eight species of genus Arcanobacterium (A.) were investigated for hemolytic properties on blood agar containing sheep and rabbit blood and for CAMP-like reactions. An enhanced hemolysis on blood agar containing rabbit blood compared to sheep blood could be observed for A. haemolyticum, less pronounced for A. hippocoleae and A. pluranimalium.A synergistic hemolytic reaction with staphylococcal β-hemolysin appeared to be constantly visible for A. hippocoleae, A. pluranimalium and A. pyogenes, with Streptococcus agalactiae for A. phocae and A. haemolyticum, with Rhodococcus equi for A. phocae, A. haemolyticum, A. pluranimalium and A. pyogenes and with A. haemolyticum for A. hippocoleae, A. pluranimalium and A. pyogenes, respectively. A reverse CAMP-reaction in the zone of staphylococcal β-hemolysin could be observed for A. phocae and A. haemolyticum. In addition, a novel CAMP-like reaction could be noted between Psychrobacter phenylpyruvicus, identified by 16S rDNA sequencing, and A. phocae and A. haemolyticum. These synergistic or antagonistic hemolytic properties could possibly be used as additional criteria for identification of bacteria of genus Arcanobacterium.     

In vitro antimycoplasmal activity of six Jordanian medicinal plants against three Mycoplasma species

The in vitro effect of six Jordanian traditional medicine plant methanolic extracts were tested against 32 isolates of Mycoplasma species; Mycoplasma mycoides subsp. mycoides LC (6), Mycoplasma capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions in Jordan. All Mycoplasma species showed susceptibility to Artemisia herba-alba and Artemisia arborescens with MIC ranges from 3.125–12.5 mg/ml. Allium sativum and Punica grantum showed limited activity against some Mycoplasma isolates. Olea europea and Citrullus colocynthis showed no in vitro activity against any of the Mycoplasma species tested. Artemisia herba-alba and Artemisia arborescens may therefore be useful for the treatment of mycoplasma infections. The authors are British Civil Servants and as such their work is subject to British Crown Copyright. This means the exclusive copyright for the article cannot be transferred.     

SNP identification, linkage and radiation hybrid mapping of the porcine lamin A/C (LMNA) gene to chromosome 4q

The lamins are components of nuclear lamina and they have a profound influence on nuclear structure and functions. They are encoded by three genes, LMNA, LMNB1 and LMNB2. A genomic fragment of the porcine LMNA gene (822 bp; from exons 7 to 9) was amplified by polymerase chain reaction and comparatively sequenced. Four single nucleotide polymorphisms (SNPs) were identified in intronic sequences: G162A, G208A, T367G and C618T. The SNPs are within the restriction sites for enzymes Bsh1236I, HpaII, AluI and Bsh1236I respectively. Allele frequencies at SNPs G208A, T367G and C618T were determined by using eight pig breeds. Linkage analysis in the Hohenheim Meishan × Piétrain family placed the LMNA gene in the chromosome 4q linkage group, between MEF2D and GBA (MEF2D– 3.0 cM –LMNA– 0.2 cM –GBA). In radiation hybrid mapping LMNA was most significantly linked to SW270 on chromosome 4 (39 cR; LOD = 7.86). The LMNA gene is located in the quantitative trait loci region for some carcass traits on chromosome 4q.     

中国伞形科当归属及相关类群毛状体和果实结构的系统学价值

深入研究了当归属及其相关7属,古当归属,高山芹属,柳叶芹属,独活属,欧当归属,山芹属及前胡属毛状体和果实结构。结果显示毛状体及果实结构(如:毛状体细胞,翅,油管,维管束,中果皮,内果皮,结合面及结晶)可以用于区分当归属及其相关类群。滇南当归及隆萼当归应从当归属移入独活属,其余当归属物种分为结合面宽及结合面窄两组,前者与前胡属较为相似,但与独活属差别较大,后者与古当归属,高山芹属,柳叶芹属,欧当归属有相似之处。支持分子系统学揭示的当归属不是个自然类群,当归属与前胡属共同位于当归分支,而独活属位于独活分支。山芹属果实结构与其他属不同,翅的中果皮无细胞,且伴生油管位于维管束内侧,为分子系统学显示此属与高山芹属,柳叶芹属,及欧当归属亲缘关系较远提供了形态学依据。     

Molecular identification and relative abundance of microorganisms in douchi koji and salted egg white sufu during processing

A novel animal protein-based douchi koji-inoculated steamed salted egg white sufu (SEWS) has been developed. This study determined the relative abundance of microorganisms in the douchi koji and semi-finished (5-day fermentation) and finished (5-day fermentation and 14-day ripening) SEWS by using 16S and 18S ribosomal DNA and gene-cloning methods. The results revealed that Bacillus spp. and Aspergillus oryzae were dominant in the douchi koji. In the semi-finished SEWS, the percentages of Bacillus amyloliquefaciens and Bacillus subtilis were considerably lower, whereas those of Enterococcus and Staphylococcus were substantially higher. In the finished SEWS, Bacillus spp. became dominant again and A. oryzae was the only fungus detected. In conclusion, by using molecular techniques, microbial population dynamics in SEWS can be evaluated. During processing, the relative abundance of microorganisms in SEWS changed and Bacillus spp. and A. oryzae remained dominant. This study provides crucial information for designing starter cultures for producing SEWS.     

Significance of β2‐Toxigenic Clostridium perfringens Infections in Animals and Their Predisposing Factors – A Review

The novel β2‐toxin of Clostridium perfringens has recently been described as the cause of enteric diseases in animals. The biological activity of β2‐toxin is similar to that of the β1‐toxin with a possibly weaker cytotoxic activity. However, the production of β2‐toxin in vitro is not seen in all β2‐toxin‐gene (cpb2)‐positive C. perfringens strains, and to deduce a clinical importance solely from the detection of cpb2 is difficult. Detection of cpb2‐positive C. perfringens from various animal species with and without enteric diseases demonstrates the wide distribution of cpb2 in nature, and the presence of cpb2 gene is therefore not considered a risk by itself. Predisposing factors like low trypsin activity in the intestinal tract, antibiotic and/or antiphlogistic treatment or changes in diet can result in the selection of β2‐toxigenic C. perfringens which may lead to enteritis or enterotoxaemia.     

The prevalence of antibodies to serovars of Leptospira interrogans in horses

SUMMARY Serum samples from 272 horses, some III, were tested by the microscopic agglutination test for the presence of antibody to 12 serovars of Leptospira interrogans. Serums from 41.5% of horses reacted to one or more of the serovars tested; the most common reactions were to L. interrogans serovar ballum (15.1%), L. interrogans serovar autumnalis (11.8%), L. interrogans serovar icterohaemorrhagiae (9.9%), L. interrogans serovar pomona (8.1%) and L. interrogans serovar hardjo (7.7%).     

Genetic diversity within two Tunisian wild jirds: Meriones shawi and Meriones libycus (Rodentia,Gerbillinae)

Three Meriones species inhabit Tunisia, namely M. shawi, M. libycus and M. crassus, but little genetic data exist on these gerbils. We collected Meriones from eight localities in Tunisia, and obtained mitochondrial (cytochrome b) and nuclear (IRBP) gene sequence data for 37 and 13 specimens, respectively, belonging to two species: M. shawi and M. libycus. We also optimised three microsatellite markers previously described in M. unguiculatus to obtain a finer analysis of their genetic diversity and geographic structure, given their wide distribution. Phylogenetic inferences of cyt b and IRBP data for these species, in the context of other gerbillin data, corroborate their taxonomic affinities reported by previous studies. High cyt b haplotype diversity was observed in both species (25 haplotypes in 29 and 27 sequences for M. shawi and M. libycus, respectively) with little geographical structure for M. shawi but three divergent groups in M. libycus. The average microsatellite diversity within each population was high (H_O ≥ 0.6, H_E ≥ 0.8) with M. libycus populations attaining the highest values. Population differentiation was moderate for several population pairs (F_st ≥ 0.1), the highest being between M. shawi populations. However, genetic distance among populations was not significantly correlated with geographic distance in either M. shawi or M. libycus. Our results contribute to a better characterisation of Tunisian Meriones species, suggesting high geographic structure in mtDNA of M. libycus populations within North Africa.     

Epidemiological Studies on Gastrointestinal Parasitic Infections of Lambs in the Coastal Savanna Regions of Ghana

The types of gastrointestinal parasites (Eimeria and helminths) encountered by 70 lambs and the seasonal pattern of both Eimeria and strongylate infections in these lambs in the derived Coastal Savanna were followed for three years. Eimeria oocysts and helminth eggs were detected in the faeces of lambs at the same time, indicating the concurrence of both Eimeria and helminth infections. Eimeria oocysts were first seen in the lambs 20 days after birth (DAB) and the level of oocyst output increased by the fourth week. Eimeria species identified in the lambs were E. parva, E. pallida, E. faurei, E. ahsata, E. bakuensis, E. intricata, E. granulosa, E. ovinoidalis and, occasionally, E. marsica. E. ovinoidalis, the most pathogenic species, dominated the oocyst output during the early part of the life of the lambs. Strongyloides papillosus eggs appeared at 46 DAB, preceding strongylate nematode eggs, which were seen at 57 DAB and those of Moniezia at 69 DAB. The pattern of Eimeria oocyst output paralleled that of the worm egg output, and high oocyst and strongylate worm egg counts corresponded with the period of high rainfall during the study period. Although oocyst and worm egg output fluctuated, high Eimeria oocyst counts were seen again in the lambs when they were 1 and 2 years old. Haemonchus species formed 71% of the infective larvae revealed by larval culture.     

An investigation into the distribution, host-preference and population density of Ixodid ticks affecting domestic animals in Bangladesh

To study the distribution, host-preference and population density of ixodid ticks in Bangladesh, an attempt was made to collect adult ticks from various host animals in three distinct topographic zones, viz. flood plains, hills and steppe ‘Barind’. Five species of ixodid ticks were recorded, namely, Boophilus microplus (56.3%), Haemaphysalis bispinosa (11.3%), Rhipicephalus sanguineus (14.7%), Hyalomma anatolicum anatolicum (15.0%) and Amblyomma testudinarium (2.8%). The data showed that B. microplus occurred predominantly on cattle (42.4%). The other hosts involved were buffaloes (12.5%), goats (25.5%) and pigs (8.2%). H. bispinosa mostly parasitized goats (31.5%) rather than cattle (12.0%) and buffaloes (10.8%). R. sanguineus was principally a dog tick (27.4%) but also parasitized cattle (10.8%) and goats (6.8%). H. a. anatolicum was restricted to cattle (19.2%) and A. testudinarium was found on both cattle (4.4%) and pigs (2.3%). These results indicate that ixodid ticks are not strictly host-specific except for H. a. anatolicum. The population density of these ticks was significantly (p < 0.01) influenced by the changing of seasons. B. microplus, H. bispinosa and R. sanguineus were by far the most widely distributed species; the distribution of H. a. anatolicum was restricted to the steppe ‘Barind tract’ and A. testudinarium was found in the hilly regions only.