and Haemophilus parasuis infection in one reaction, through a melting curve analysis. This method utilized two pairs of specific primers
that allowed the amplification of highly conserved regions of A. pleuropneumoniae Apx IV and H. parasuis omp P2 gene. Reconstitution
experiments were conducted by using PMD - 19T plasmid in order to determine the sensitivity of the assay. The results showed that the Tm
values of the melting curves of A. pleuropneumoniae and H. parasuis were 83.36±0.09ºC and 76.48±0.17ºC, respectively that could accurately
distinguish these two pathogens. And no cross reaction were observed between other respiratory pathogens, which suggested a high
specificity of two primers. The detection sensitivity of the assay was 127 and 96 copies/μL which was higher than that of the ordinary PCR
detection methods. This rapid technique may present a simple, useful option for simultaneous detection of A. pleuropneumoniae and H.
parasuis, which would be feasible and attractive for clinical samples diagnosis and epidemiological investigations.
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Ling PENG , Xiaoqing YUAN, Ran FANG, Weizhen LIU-FU, Quan WEN, Xufu YANG
Ling PENG ,
Xiaoqing YUAN, Ran FANG, Weizhen LIU-FU, Quan WEN, Xufu YANG
This study analyzed the characteristics of 133 Haemophilus parasuis isolates in southern China. These isolates belonged to eleven serovars (1, 2, 4-10, 13 and 15) with 12.0% of them being characterised as non-typable. A relatively high level in resistance was encountered for trimethoprim + sulfamethoxazole (89.9%), tetracycline (75.3%), amoxicillin (69.1%), streptomycin (63.6%), carbenicillin (60.2%), kanamycin (46.6%) and ampicillin (45.6%). A total of 60% of the isolates were negative for group 1 virulence-associated autotransporters (vtaA). All group 1 vtaA negative isolates fell into polyacrylamide gel electrophoresis (PAGE) type I, while all group 1 vtaA positive isolates were classified as PAGE type II. The results of Multi-locus sequence typing (MLST) indicated a high degree of variation, 45 isolates in the study were assigned into 31 sequence types with 28 of these being new (not found in the MLST database). Antimicrobial resistance was observed in every serovar, there was no statistically significant correlation between the antimicrobial resistance and the serovars. The isolates allocated to clade 2 (based on MLST target sequences) showed the molecular characteristics of highly pathogenic strains in whole-cell protein profiling, vtaA groups 1, superoxide dismutase (sodA) sequence and MLST.
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Alkyd Resin method, patented by the Turkish Patent Institute, is a cadaver preparation and preservation technique. This study examines whether exhibition-museum materials can be produced from rat cadavers processed with alkyd resin method in different postures and whether dissection and suturing can be performed on the samples. Besides, samples prepared with alkyd resin and samples prepared freshly were compared in terms of dissection and suturing. 14 Wistar Albino Rats were used in the study. Seven were prepared with alkyd resin method and the others were freshly prepared for comparison in terms of suturing and dissection. The alkyd resin method was applied to two cadavers as a pretrial and to five cadavers for them to be museum-exhibition and educational materials. Alopecia was detected in two cadavers. In some of the exhibition samples processed with the method mentioned, shrinkage and hardening of the skin was noticed. All the samples prepared with this method became products suitable for suturing and dissection. The exhibition-museum materials prepared with the alkyd resin method have maintained their first body postures. We believe that alkyd resin method can be applied to the entire body, and the products can be used for exhibition and educational purposes. Furthermore, the method should be further popularized, and it should be applied to cadavers of different species so that its applicability increases.
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The aim of this study was to investigate the effects of two modified semen diluents on the spermatological parameters of Saanen goat sperm stored at 4°C. Ejaculates were obtained from five Saanen bucks by means of an electro-ejaculator and divided into five aliquots. Then, aliquots of ejaculates were used to create five experimental groups as follows: Group 1 (control), group 2A containing seminal plasma (SP) and bovine serum albumin (BSA; 10 mg/mL-1), group 2B containing BSA without SP, group 3A containing egg yolk (EY; 17.00%) and SP and finally group 3B containing EY without SP. All aliquots of semen were extended with Tris-based extender. Spermatological parameters of experimental groups were observed at 0, 6, 12 and 24 h periods during storage at 4°C. Motility of semen in groups 2A and 2B was found to be best values compared to other groups. However, the motility of group 3A at 24 h was 0.00%. In conclusion, it was concluded that the addition of 10 mg/mL-1 of BSA to Tris-based extender could be useful in the storage of Saanen goat semen regardless of SP at 4°C.
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Transmissible venereal tumor (TVT) is a transmissible cancer that typically affects the external genital organs in canines. It can spread to other parts of the body via auto- and hetero-transplantation in a way that neoplastic cells are inoculated into the degraded mucosa or skin during the interaction of the animals, such as licking and sniffing, or less commonly via hematogenous or lymphatic routes. Here, we presented the clinical, cytological, and histopathological characteristics of two rare cases of TVT that developed ocular involvement and were associated with a poor prognosis. Fine needle aspiration biopsy was the initial method of choice in the diagnosis which is rapid, minimally invasive, and effective. It was diagnostic in one case and aided in the diagnosis in the other. The latter was considered a malignant round cell tumor in the cytological examination that needed further clinical and histopathological examination for definitive diagnosis. TVT is a round cell tumor that should be considered in the differential diagnosis of ocular lesions in canines.
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Hilal ACAR ,
Ece ÇERÇİ ,
Marzieh Karimi KHEZRI ,
Melike ÇETİN ,
Uygur CANATAN ,
Mehmet Metin ŞEN ,
Vildan ASLAN ,
Canan ALTINCI SARIL ,
Elif MEKİK TEMİZ ,
MEVLÜT ÖZGÜR TAŞKAPILIOĞLU ,
Hatice ERDOST ,
Hakan SALCI
In this study, histologic and electromyographic (EMG) evaluation of neuroregenerative effect of stromal vascular fraction (SVF) following periferal nerve anastomosis was aimed. Totally, 31 Wistar Albino, male rats (weighing about 300 g) were studied, and these rats were grouped according to surgical techniques performed on the right sciatic nerve: group I (GRI) micro suture (n=7), group II (GRII) micro suture + SVF (n=7), group III (GRIII) fibrin glue (n=7) and group IV (GRIV) fibrin glue + SVF (n=7). Fat tissue was taken from 3 rats to prepare SVF, and SVF was produced by non-enzymatic method. The left sciatic nerve of all rats was evaluated for EMG as control. Under general anesthesia, after transversal incision of the sciatic nerve, microsurgical epineural repair technique was performed with 10/0 nonabsorbable suture. EMG examinations were performed in terms of conduction rate, amplitude, distal latency and spontaneous muscle activity at 0th day and postoperative (PO) 1st and 8th weeks. EMG results between and within the groups were statistically evaluated by one-way analysis of variance. Rats in all groups were sacrificed by decapitation at PO 8th week and histological examinations of the sciatic nerves were performed following preparation of the neural tissues. EMG examination results showed the highest nerve conduction in GRI, the highest amplitude in GRIII, normal latency in GRII and longer distal latency in GRIV at PO 8th week. Amplitude and conduction velocity increased gradually in all groups. In needle EMG, the best muscle membrane stabilization was achieved in GRII and GRIV at PO 8th week. Statistically, the values of amplitude, distal latency, conduction velocity, and spontaneous muscle activity were found to be at normal levels at PO 8th week in all groups (P>0.05). In the histological results, although fibroconnective tissue reactions in the anastomosis area had similar scores in GRII and GRIV, maximum fibroconnective tissue reaction and the best axonal regeneration was seen in GRI and GRIII, respectively. In addition, GRI and GRII had the most inflammatory cells accumulation in the suture region, and less inflammatory cells were seen in the anastomosis area of GRIII and GRIV. As a conclusion, fibrin glue presents good electrophysiological and histological results; however, it is clear that local SVF usage on the nerve anastomosis area can be a good choice to decrease fibroconnective tissue reaction and inflammation.
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Xingxing ZHANG ,
Jie LI ,
Jun QIAO ,
Qingling MENG ,
Ling ZHANG ,
Ying CHEN ,
Kuojun CAI ,
Xin HUANG ,
Tongzhong WU ,
Tongzhong WU ,
Fagang ZHONG
The widespread multidrug-resistant Escherichia coli strains have caused a severe challenge to animal health and the development of breeding
industries. The purpose of this study was to investigate the phylogenetic grouping and antimicrobial resistance profiles of E. coli isolated from
diarrheic calves in Xinjiang province, China. In this study, a total of 379 E. coli strains were isolated from 379 rectal swab samples of diarrheic calves.
They were further analyzed their phylogenetic groupings by multiplex PCR, and were clustered into four phylogenetic groups, A (36.1%), B1
(17.4%), B2 (15.6%), and D (30.9%). All E. coli isolates were tested for their susceptibility to 15 antimicrobial agents by Kirby-Bauer (KB) method. The
isolates showed the highest resistance rates against ampicillin (64.9%), followed by streptomycin (59.4%), tetracycline (53.8%), sulfamethoxazole/
trimethoprim (50.9%), chloramphenicol (45.6%), kanamycin (44.1%) and enrofloxacin (42.0%). E. coli isolates exhibited lower resistance to
ceftazidime (15.0%) and polymyxin (12.6%). The resistance genes blaTEM, blaOXA, mcr-1, strA-strB, aadA, tet(A), tet(B), and tet(C) were detected in
68.3% (168/246), 27.2% (67/246), 14.6% (7/48), 51.1% (115/225), 24.9% (56/225), 51.5% (105/204), 44.6% (91/204), and 7.8% (16/204) of E. coli
isolates, respectively. These results demonstrate that prevalent multi-drug resistance and high level of antimicrobial resistance genes exist among
E. coli from Xinjiang diarrheic calves and pose a potential public health concern
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Mycoplasma mastitis raises significant concerns in the dairy industry worldwide. The study objective was to develop an accurate and rapid
screening method for identification of field isolates of Mycoplasma and Acholeplasma species and investigate relative merits of conventional
microbial culture versus PCR-based method for detecting Mycoplasma and Acholeplasma species in bovine milk. A total of 368 milk samples
collected at individual cow level from a single dairy farm in South Australia, 192 (52%) tested positive for mollicutes using a conventional culturebased method. DNA extracted directly from milk and used for amplification through specifically designed universal mollicutes PCR-based
method. Of them, 269 (73%) tested positive. Sequencing results of 30 positive samples targeting the 16S rRNA gene, showed five different
mollicutes species involved, including Acholeplasma laidlawii, Acholeplasma axanthum, Mycoplasma arginini, Mycoplasma bovirhinis, Mycoplasma
bovis. According to these results, species-specific PCR was conducted on all samples. DNA amplifications using species-specific PCR yielded
256 (70%) positive mollicutes samples. The developed universal PCR demonstrated best concordance with species-specific PCR (Cohen’s Kappa
= 0.747±0.031). Co-infection by two or more of the above-mentioned mollicutes showed highest prevalence. It is recommended surveying
mollicutes using the universal PCR used in this study. The PCR system used in this study showed significant rapidity and sensitivity compared to
the conventional bacteriological culture method for screening Mycoplasma and Acholeplasma species in dairy herds.
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