Bacterial communities in buffalo rumen were characterized using a culture-independent approach for a pooled sample of rumen fluid from 3 adult Surti buffaloes. Buffalo rumen is likely to include species of various bacterial phyla, so 16S rDNA sequences were amplified and cloned from the sample. A total of 191 clones were sequenced and similarities to known 16S rDNA sequences were examined. About 62.82% sequences (120 clones) had >90% similarity to the 16S rDNA database sequences. Furthermore, about 34.03% of the sequences (65 clones) were 85-89% similar to 16S rDNA database sequences. For the remaining 3.14%, the similarity was lower than 85%. Phylogenetic analyses were also used to infer the makeup of bacterial communities in the rumen of Surti buffalo. As a result, we distinguished 42 operational taxonomie units (OTUs) based on unique 16S r DNA sequences: 19 OTUs affiliated to an unidentified group (45.23% of total OTUs), 11 OTUs of the phylum Firmicutes, also known as the low G+C group (26.19%), 7 OTUs of the Cytophaga-Flexibacter-Bacteroides phylum (16.66%), 4 OTUs of Spirochaetes (9.52%), and 1 OTU of Actmobacteria (2.38%). These include 10 single-clone OTUs, so Good's coverage (94.76%) of 16S rRNA libraries indicated that sequences identified in the libraries represent the majority of bacterial diversity present in rumen.
In the milk industry in India, buffalo breeds are most commonly used for milk production. Efficiency of fiber digestion in ruminants is critical for animal productivity. Bacteria play an important role in fiber digestion and utilization. Absolute quantification real-time PCR was used to quantify ten bacterial species in rumen fluid of Surti buffalo fed green fodder, dry roughage and compound concentrate mixture. Abundance of each target taxon was calculated as a fraction of the total 16S rRNA gene copies in the samples, using taxon-specific primers. Bacterial populations showed a clear predominance of Ruminococcus albus, which comprised 5.66% of the bacterial rRNA gene copies in the samples. However, only 0.9% to 4.24% of the bacterial rRNA gene copies were represented by the ruminal Fibrobacter succinogenes, Ruminococcus flavefaciens and Prevotella species. The proportion of rRNA gene copies attributable to Selenomonas ruminantium, Streptococcus bovis, Ruminobacter amylophilus, Treponema bryantii and Anaerovibrio lipolytica was even less abundant, each comprising <0.11% of the bacterial rRNA gene copies. The data suggest that the aggregate abundance of the most intensively studied ruminal bacterial species is relatively low and that a large fraction of the uncultured population represents a single bacterial genus.
Fasciolosis caused by Fasciola spp. is considered the most important helminth infection of ruminants in tropical countries. Anthelmintic resistance has become a global concern. This study compared the efficacy of the commonly used anthelmintics, determined the toxicity level and any indication of resistance. Thirty two water buffaloes naturally-infected with Fasciola spp. were used to determine the efficacy of triclabendazole (TBZ), albendazole (ABZ), and bromofenofos (BRO) using Fecal Egg Count Reduction Test (FECRT). To test the toxicity of the drugs given, serum glutamic-pyruvic transaminase (SGPT) was evaluated before and within one week after treatment. One dose administration of ABZ registered an efficacy of 79.17%, 73.33% for TBZ and 70.83% for BRO. Efficacy in two dosetreatment group was 83.33% for both BRO and ABZ, and 90.00% for TBZ. Two dose-treatment was effective for TBZ (90%), ineffective for BRO and ABZ. SGPT levels were not significantly different between pre-treatment and posttreatment across all treatments. Giving one or two doses of anthelmintics, at one month interval, does not increase the efficacy of the three drugs tested. The study also implies that anthelmintic resistance may have developed in the animals.
The objective of the study was to compare the usefulness of FLOTAC and centrifugal fecal flotation (CFF) techniques. More specifically, the taxonomic classes (Nematoda and Cestoda) of endoparasites present in fecal samples of buffaloes are identified, the sensitivity and specificity of FLOTAC relative to CFF are calculated, and the agreement of both techniques is evaluated using Kappa statistics. Fresh fecal samples from 220 buffaloes in 10 municipalities were collected. Sheather’s sugar was used as a flotation solution for both the FLOTAC and CFF techniques. Of the 220 animals, 109 samples were nematode positive and 111 samples were nematode negative according to the FLOTAC technique, while 74 were found to be positive and 146 negative according to the CFF technique. No cestodes were detected by either technique. The calculated sensitivity for FLOTAC is 89.19% and its specificity is 70.55%. Kappa statistics revealed moderate agreement (k=0.535) between the two techniques in detecting nematodes. The prevalence observed based on FLOTAC and CFF test were 49.54% (109/220; 95% CI: 47.75–56.34) and 33.64% (72/220; 95% CI: 27.42–40.3), respectively.
Trypanosoma evansi infection in the Philippines is frequently reported to affect the country’s livestock, particularly, the buffaloes. To assess the prevalence and intraspecific diversity of T. evansi in the country, blood samples from water buffaloes in different geographical regions were collected during an outbreak. T. evansi was detected in all 79 animals tested using PCR targeting the RoTat 1.2 VSG gene. Sequencing of the rDNA complete internal transcribed spacer (ITS) region including the 5.8S subunit showed high similarity (99–100%) between Philippine isolates and known T. evansi isolates in Genbank. Tree construction based on the same region confirmed the close relationship between Philippine and reported Thai isolates as compared to Egyptian isolates separated by relatively small genetic distances, 47 polymorphisms, despite the clustering in four branches. Overall, the results of this study prove genetic diversity within T. evansi species despite previous reports on limited heterogeneity among isolates worldwide.
CD14 plays a crucial role in the inflammatory response to lipopolysaccharide (LPS), which interacts with TLR4 and MD-2 to enable cell activation, resulting in inflammation. Upstream inhibition of the inflammation pathway mediated by bacterial LPS, toll-like receptor 4 (TLR4) and cluster of differentiation antigen 14 (CD14) was proven to be an effective therapeutic approach for attenuating harmful immune activation. To explore the effect of CD14 downregulation on the expression of TLR4 signaling pathway-related genes after LPS stimulation in buffalo (Bubalus bubalis) monocyte/macrophages, effective CD14 shRNA sequences were screened using qRT-PCR and FACS analysis with buffalo CD14 shRNA lentiviral recombinant plasmids (pSicoRGFP-shRNA) and buffalo CD14 fusion expression plasmids (pDsRed-N1-buffalo CD14) co-transfected into HEK293T cells via liposomes. Of the tested shRNAs, shRNA-1041 revealed the highest knockdown efficiency (p < 0.01). When buffalo peripheral blood monocyte/macrophages were infected with shRNA-1041 lentivirus and stimulated with LPS, the expression of endogenous CD14 was significantly decreased by CD14 shRNA (p < 0.01), and the mRNA expression levels of TLR4, IL-6 and TNF-α were also significantly downregulated compared to the control groups (p < 0.01). These results demonstrated that the knockdown of endogenous CD14 had clear regulatory effects on the signal transduction of TLR4 after stimulation with LPS. These results may provide a better understanding of the molecular mechanisms of CD14 regulation in the development of several buffalo diseases.
To investigate the prevalence of Neospora canimim antibodies in water buffalo (Bubalus bubalis) in the north-western region of Iran, blood samples were taken from 181 (141 females and 40 males) buffaloes slaughtered in an abattoir in Ahvaz. Sera were tested for the presence of antibodies to N. canimim by an ELISA kit. The overall prevalence of Neospora canimim infection in the buffaloes tested was 37% (43.36% in females and 15% in males). Statistical analysis showed significant differences between male and female buffaloes. In female buffaloes, 7.7% of heifers and 56.8% of cows were seropositive. Statistical analysis showed the differences among female buffaloes were highly significant when their age is considered, and buffalo cows showed higher N. caninum prevalence when compared with heifers. With regard to the results of this study, high exposure of water buffaloes to N. canimim, and economical importance of buffalo industry to some regions of Iran, the role of N. canimim as a causal agent of abortions in this species should be investigated.
This study was conducted to compare the time of onset, duration of action, and the extent of analgesia produced by a lidocaine/xylazine combination with those produced by lidocaine and xylazine alone after injection into the caudal extradural space of the Iranian river buffalo. The study was designed as a prospective, descriptive, observer-blind trial, in a Latin square pattern. Eleven adult (aged over 2 years) non- gravid and healthy females of Iranian river buffaloes (Bubalus bubalis), weighing from 450 to 650 kg, were used. Caudal extradural analgesia was achieved on 3 occasions at 14 d intervals by injection of 2% lidocaine (L; 0.22 mg kg⁻¹), 2% xylazine (X; 0.05 mg kg⁻¹), and a combination of 2% lidocaine(0.22 mg kg⁻¹) / 2% xylazine (LX; 0.05 mg kg⁻¹) in a Latin square design. Analgesia was determined by the lack of response to pinprick and haemostat pressure in the skin of the caudal areas. X was significantly longer (5.5 ± 0.7 min) than that by L or LX. Duration of analgesia was significantly longer by LX (172.3 ± 17.7 min) than that by either drug used alone (lidocaine, 79.5 ± 5.7 min; xylazine, 136.4±11.4 min). In X and LX groups, the level of analgesia ascended to thoracic segments; however, in lidocaine-treated buffaloes thighs, flank, and udders remained sensitive. In all the buffaloes, xylazine, administered either alone or with lidocaine, induced mild to moderate ataxia. It was concluded that the LX combination provided a more rapid onset and a longer duration of analgesia, and a more cranial spread of analgesic effect compared with either drug alone. As a result, the LX combination may offer a fast and long lasting anaesthesia/analgesia to perform obstetrical and surgical procedures without the need for re-injection.
Concentration of selenium in serum samples was determined using a modified Watkinson's spectrofluorometric method. Selenium concentration averaged 0.021 ±0.008 µg/mL for water buffaloes and 0.074 ±0.017 µg/mL for dairy cows. The analysis of selenium concentrations in cows showed deficiency level in 2.0%, marginal level in 64%, and optimal level in 34% of the examined samples. In water buffaloes, deficiency level was found in 96.55% and marginal level in 3.45% of the animals. Selenium levels in cattle and water buffaloes should be periodically diagnosed to provide data for veterinarians to make appropriate decisions. The implementation of suitable prophylactic programmes will enable optimum levels of this element to be maintained.