Epidemiological survey on <Emphasis Type="Italic">Escherichia coli</Emphasis> O157 in Chongqing and Three-Gorge Reservoir Areas of China

Prevalence, presence of virulence and adherence associated genes, genetic diversity, biochemical characteristics, and antibiotic susceptibility were determined for Escherichia coli O157 isolated over 4 months in Chongqing city and Three-Gorge Reservoir Areas. 11 isolates of E. coli O157 were isolated from 1504 samples and 7 of them are O157:H7 and 4 are O157:H? All O157:H7 isolates had eaeA, ehxA, EspA and Tccp genes, but did not have stx1 and stx2. All O157:H? isolates did not have stx1, stx2, eaeA, ehxA, EspA and Tccp genes except for the isolate obtained from Yunyang county which had stx1. When eaeA and ehxA presented in isolates were digested by restriction enzymes, the numbers and the sizes of the segments were the same as the control E. coli O157 strains. This suggests that eaeA and ehxA exhibit poor polymorphism. Most E. coli O157 isolates showed identical biochemical activities to the standard strains for sorbitol and rhamnose, and all E. coli O157:H7 obtained from feces at the same dairy cattle farm had similar biochemical characteristics. Antibiotic susceptibility demonstrated resistance of the isolates to penicillin, ampicillin, bacitracin, cefuroxime, erythromycin, gentamycin and tetracycline, indicating the isolates obtained in this study had a multi-drug resistance.     

Polymorphism of growth hormone receptor (<Emphasis Type="Italic">GHR</Emphasis>) gene in Nilagiri sheep

The allelic variation in the regulatory sequence of growth hormone receptor (GHR) gene influences the growth traits of sheep. A study was carried out to find out the polymorphisms associated with exon 10 of GHR gene and its association with growth traits of Nilagiri sheep. The blood samples were collected from Nilagiri sheep (n?=?103) reared at Sheep Breeding Research Station, Sandynallah, Tamil Nadu, India. DNA was isolated using the phenol-chloroform extraction procedure and eight samples having amplified product of part of exon 10 (895 bp) sequenced. The results indicated transitions of nucleotide G>A at loci G177624A and G177878A. The genotyping frequencies estimated using the tetra-primer amplification refractory mutation system-PCR for GG, GA and AA were 0.262, 0.544 and 0.194, and 0.349, 0.505 and 0.146, respectively. The estimated allele frequencies of G and A nucleotides were 0.5340 and 0.4660, and 0.6015 and 0.3985, respectively, at loci G177624A and G177878A. The effects of both the mutations on growth-related traits viz., birth, weaning (3 months) 6, 9 and 12 months weight in Nilagiri sheep were found to be non-significant. This can be a novel approach to assess growth of sheep using the mutation in GHR gene. Thus, this approach can be useful for further investigation as a molecular marker associated with genetic improvement.     

Serological survey for <Emphasis Type="Italic">Brucella</Emphasis> antibodies in donkeys of north-eastern Nigeria

A cross-sectional epidemiological study was conducted to determine seroprevalence and risk factors influencing the presence of Brucella antibodies in donkeys of Borno State, north-eastern Nigeria. The study aimed at providing baseline information that may be used in planning a control policy against equine brucellosis. Blood samples were collected from 601 donkeys, comprised of 374 males and 227 females from the six agricultural zones of the state between March 2013 and September 2014. The sera obtained were tested for Brucella antibodies using Rose Bengal plate test (RBPT) and competitive enzyme-linked immunosorbent assay (cELISA). Of the 601 donkeys tested, 43 (7.2%) and 40 (6.7%) were seropositive by RBPT and cELISA, respectively. A seroprevalence of 8.6% was obtained in male and 3.5% in female donkeys. According to age, the highest seroprevalence of 9.6% was obtained from donkeys of age group 4–6 years. With respect to pregnancy status, a higher seroprevalence (6.8%) was obtained from pregnant donkeys compared to 3.8% obtained from the non-pregnant ones. There were statistically significant associations between the presence of antibodies and sex (p < 0.05) and the presence of antibodies and age (p < 0.05) of the studied donkeys. However, no statistically significant association (p > 0.05) was observed between the pregnancy status and presence of antibodies. The study concludes that Brucella infection is present in donkeys in all the agricultural zones of the state. The relatively high seroprevalence (7.2%) obtained is of public health concern because of the close interaction between donkeys, ruminants, and humans in the study area.     

The combined effects of the provision of feed and healthcare on nutrient utilization and growth performance of sheep during the early or late dry season

An on-farm study was conducted to determine the combined effects of the provision of feed and healthcare on nutrient use and growth performance of sheep during the early or late dry season. A total of 36 smallholder sheep farmers with a flock size of ≤7 was randomly selected within each of the three administrative regions in Northern Ghana. The sheep grazed on a heterogeneous natural pasture and offered crop residues as basal diet (control) or were additionally provided with a concentrate feed plus orthodox healthcare to control diseases and pests (CH) in a completely randomized block design. The provision of orthodox healthcare included scheduled control of endo- and ecto-parasites and administration of broad-spectrum antibiotics. Data was analyzed for the fixed effects of CH, season, or CH × season using the mixed model procedure of Genstats^®. The CH regimen had no effect (P = 0.098) on intake of natural pasture but pasture intake increased (P = 0.012) during the late dry season. Sheep on the CH regime had higher DM (P = 0.026) and N (P = 0.068) digestibility and improved ADG (P = 0.001) and feed conversion efficiency (P = 0.020) than those on the control. We hypothesize that improvements in growth performance of sheep on the CH regimen could be related to availability of nutrients for growth that will otherwise have been used for repair of damaged tissues caused by gastrointestinal parasites and ticks. Sheep on the CH regimen also had a higher concentration of fecal N during the late dry season when CP concentration was relatively higher than that in the early dry season (63.2 vs 60.9 g/day DM) when CP concentration of pasture was lower.     

Genotypic and phenotypic detection of capsular polysaccharide and biofilm formation in <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolated from bovine milk collected from Brazilian dairy farms

Staphylococcus aureus is a pathogen that frequently causes mastitis in bovine herds worldwide. This pathogen produces several virulence factors, including cell-associated adhesins, toxic and cytolytic exoproteins, and capsular polysaccharides. The aim of the present study was to test for the presence of genes involved in capsular polysaccharide production and biofilm formation in S. aureus isolated from bovine mastitis samples collected from 119 dairy herds located in three different Brazilian regions, as well as to assay the production of capsular polysaccharides and biofilm, in vitro. The detection of the cap, icaAD, and bap genes was performed using PCR. The detection and quantification of capsular polysaccharide production was performed using ELISA assays. The ability of the isolates to form a biofilm was examined using the polystyrene surface of microtiter plates. All 159 S. aureus isolates investigated harboured the cap gene: 80 % carried the cap5 gene and 20 % carried the cap8 gene. Sixty-nine percent of the isolates expressed capsular polysaccharide (CP) in vitro, 58 % expressed CP5 and 11 % expressed CP8. All of the isolates harboured the icaA and icaD genes, and 95.6 % of the isolates carried the bap gene. Of the 159 isolates analysed, 97.5 % were biofilm producers. A significant association between the capsular genotype and phenotype and the amount of biofilm formation was detected: cap5/CP5 isolates tended to form more biofilm and to produce a thinner CP layer than cap8/CP8 isolates. The results indicate a high potential for pathogenicity among S. aureus isolated from bovine milk collected from three different regions in Brazil.     

Physio-biochemical parameters: a potential tool for target-selective treatment of haemonchosis in the small ruminants

This study aims to evaluate the conjunctiva colour-based FAMACHA score (FS) coupled with a body condition score (BCS), haemogram and stressor hormone level estimation, in identifying post-mortem (PM)/coproscopically proven individuals wanting therapy for economically important gastrointestinal (GI) helminths, Haemonchus contortus, in the small ruminants. The incidence of haemonchosis was significantly (p < 0.05) higher (60.81%) in the ruminants with FS = 3. The H. contortus count in the animals with FS 2, 3 and 4 was 23.2 ± 0.37, 62 ± 2.5 and 74 ± 3.2 (p < 0.05) [positive correlation (r = 0.841 in goats; r = 0.828 in sheep, p < 0.05)], respectively, with corresponding 2.8 ± 0.15, 2 ± 0.3 and 2 ± 0.16 BCS (negative correlation, p > 0.05). The infected animals of FS 2, 3 and 4 measured 8.2 ± 0.0, 7.5 ± 0.23 and 6.7 ± 0.34 g/dl Hb (r = ?0.452, p = 0.01) in goats/9.3 ± 0.8, 8.6 ± 0.5 and 7.6 ± 0.3 g/dl Hb (r = ?0.511, p = 0.05) in sheep with 21.2, 19.8 ± 1.8 and 17.8 ± 0.2% PCV (r = ?0.369, p = 0.05) in goats/26.7 ± 1.2, 22.2 ± 0.2 and 20.9 ± 0.6% PCV (r = ?0.251, p = 0.03) in sheep, respectively. The FS 2, 3 and 4 infected goats/sheep measured 6.1 ± 0, 7.9 ± 1.0 and 9.5 ± 0.9 (p < 0.05)/5.8 ± 2.3, 6.9 ± 1.2 and 7.8 ± 0.2% (p < 0.05) mid-granulocyte [(r = 0.928 (goats)/0.834 (sheep), p < 0.05], while the cortisol level was 15.6, 23 ± 4.5 and 42 ± 2.3 (p = 0.23)/12.1 ± 0, 15.9 ± 1.2 and 24 ± 3.4 (p = 0.29) μg/dl, respectively. The infected ruminants recorded low (p < 0.05) level of Hb/PCV while high level of mid-granulocytes/cortisol. Specificity of FAMACHA test was maximized (100%) when FS = 4 was considered anaemic, but sensitivity was low (35.29% in goats; 25% in sheep). The false negatives was 5.9 (goat)/12.5 (sheep)% when FS ≥ 3 was considered anaemic. The small ruminants with FS ≥ 3, BCS ≤ 2.5, Hb ≤ 7.5 g/dl (goats)/8.6 g/dl (sheep), PCV ≤ 19.8% (goats)/22.2% (sheep) and mid-granulocyte ≥7.9% (goats)/6.9 ± 1.2% (sheep) can be subjected to target-selective treatment for haemonchosis in the field simultaneously maximizing the economic benefit to the farmers.     

In vitro control of parasitic nematodes of small ruminants using some plant species containing flavonoids

This study determined in vitro anthelmintic efficacy of three plant species: Trema orientalis, Urtica dioica and Zanthozylum capense on nematode larvae of small ruminants. Dried leaf samples (40 g) were extracted in 70% ethanol, in portions of 10 g and concentrated to 100 ml. Half and one quarter of the original crude extract were both made to 100 ml. Rectal faecal material from 10 Merino sheep and 25 Nguni goats was pooled within species and thoroughly hand-mixed. Dung samples, each of 5 g were cultured for 12 days at 27 °C. On day 13, 4 plates were watered and 4 others treated with ethanol to correct for solvent effect on mortality. The design was 2 (animal species)?×?3 (plant species)?×?3 (extract concentrations). In each of three runs, three plates were treated with each crude extract in three incremental concentrations. Surviving L3 larvae were isolated, counted and mortalities became indices of anthelmintic efficacy. Data from nematode larval mortality were analysed to determine the effect of animal species, plant species, concentration and their interactions. Efficacy was affected by concentration (P?=?0.0001), animal species (P?=?0.0046), plant species (P?=?0.0572), the interactions of animal species and concentration (P?=?0.0010), plant species and concentration (P?=?0.0123) and concentration × animal × plant species (P?=?0.0435).     

SCC <Emphasis Type="Italic">mec</Emphasis> typing and antimicrobial resistance of methicillin-resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> (MRSA) from pigs of Northeast India

Staphylococcus aureus is one of the most important pathogens of both humans and animal. Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen that causes serious infections both in hospitals and communities due to its multidrug resistance tendency. This study was undertaken to characterize the MRSA isolates from pigs and to determine the antimicrobial resistance of these isolates. Forty nine MRSA strains (one strain per positive pig) isolated from pigs of Northeast India were characterized by SCCmec typing and antimicrobial resistance. The overall prevalence of MRSA was 7.02 % with the highest prevalence recorded in pigs aged 1–3 months (P = 0.001) and in nasal samples (P = 0.005). Two SCC mec types (type III and V) were found in Indian pigs with predominance of type V. All isolates were resistant to penicillin. Seventeen resistance groups were observed where 87.75 % isolates showed multidrug resistance (showed resistance to three or more classes of antimicrobials). The most predominant resistance pattern observed was Oxytetracycline + Penicillin + Sulfadiazine + Tetracycline accounting 12.24 % of the isolates. The present study contributes to the understanding of characteristics and antimicrobial resistance of porcine MRSA isolates which in turn will help in devising strategy for the control of this pathogen. Findings of the study also throw light on multidrug resistance MRSA and emphasize the need for judicious use of antimicrobials in animal practice.     

Cryptosporidium species detected in calves and cattle in Dagoretti,Nairobi, Kenya

A total of 1,734 cattle faecal samples from 296 dairy-keeping households were collected from urban settings in Nairobi, Kenya. Modified Ziehl–Neelsen staining method and an immunofluorescence assay were used to identify those samples with Cryptosporidium oocyst infection. Oocysts from positive faecal samples were isolated by Sheather's sucrose flotation method and picked from the concentrate using cover slips. Genomic DNA was extracted from 124 of the faecal samples that were positive for Cryptosporidium and was used as template for nested PCR of the 18S rRNA gene. Twenty-five samples (20 %) were PCR-positive for Cryptosporidium, and 24 of the PCR products were successfully cloned and sequenced. Sequence and phylogenetic analysis identified 17 samples (68 %) as Cryptosporidium parvum-like, four samples (16 %) as Cryptosporidium ryanae, three samples (12 %) as Cryptosporidium andersoni and one sample (4 %) as Cryptosporidium hominis. To the best of our knowledge, this is the first genotyping study to report C. parvum-like, C. andersoni and C. hominis in cattle from Kenya. The results of this study show Cryptosporidium infections in calves and cattle may be potential zoonotic reservoirs of the parasite that infects humans.     

Prevalence of <Emphasis Type="Italic">Haemonchus contortus</Emphasis> infection in sheep slaughtered at Jimma town municipal abattoir,Ethiopia

Haemonchus contortus is a parasite of major economic importance of most sheep-rearing areas of the world. A cross-sectional study was conducted from November 2015 to March 2016 with objectives of determining the prevalence and associated risk factors of Haemonchus contortus infection in sheep slaughtered at Jimma town municipal abattoir, Ethiopia. Of the total 7000 sheep slaughtered during the study time period, 384 sheep were randomly selected and examined for parasites. The overall prevalence of Haemonchus contortus infection was 264 (68.75%). Infection rate of poor body condition animals was significantly (P?=?0.000) higher than good body conditioned animals. The mean packed cell volume (PCV) values (%) of parasitemic and aparasitemic sheep were 23.73?±?3.7 SD and 37.66?±?4.736 SD, respectively. The prevalence in adult sheep (>?1 year) was insignificantly (P?=?0.653) higher than that of young sheep (≤?1 year). The highest prevalence was recorded in sheep that originated from Seka district (73.3%) and the lowest in Kersa district (63.4%) with non-significant variation (P?=?0.691). The highest monthly mean worm burdens and prevalence of Haemonchus contortus infection were recorded in November and lowest in March. The current study revealed that Haemonchus contortus infection is an important and common parasitic disease and requires special attention to its control.     

Role of Chlamydophila abortus in ovine and caprine abortion in Sardinia,Italy

Between 1999–2003, 14 321 sera and 646 abortion samples (498 foetuses and 148 placentae) were analysed from 807 sheep and goat farms distributed all over the island of Sardinia. After notification of abortion in a flock, sera collected at random from adult animals were examined to detect antibodies specific to Chlamydophila (C.) abortus by ELISA, whereas foetuses and placenta were analysed by PCR assay. Specific IgG antibodies were detected in 611 (4.8%) sheep and 106 (5.8%) goats. From a total of 2050 ovine and 151 caprine fetal samples including muscle, liver, abomasum, spleen, brain and placenta, 29 (1.4%) ovine and 1 (0.6%) caprine samples were C. abortus PCR-positive. Placenta was the tissue with the highest detection rate. These results indicate that the seroprevalence of C. abortus infection in sheep and goats is very low in Sardinia, and PCR results demonstrate that C. abortus has no significant role in abortion, especially in goats.     

Immunoprotection of recombinant <Emphasis Type="Italic">Eg</Emphasis>.P29 against <Emphasis Type="Italic">Echinococcus granulosus</Emphasis> in sheep

Objective

This study aims to investigate the immunoprotection of recombinant Eg.P29 (rEg.P29) vaccine and analyze the underlying mechanism in sheep.

Methods

Three groups of male sheep were immunized subcutaneously with rEg.P29 and PBS, Freund’s complete adjuvant as controls, respectively. After prime-boost vaccination, the sheep were challenged with encapsulated Echinococcus granulosus eggs. The percentage of protection in sheep was determined 36 weeks after the infection. Humoral immune response was analyzed for specific IgG, IgG1, IgG2, IgM and IgE levels. Moreover, cytokines including interferon (IFN)-γ, interleukin (IL)-2, IL-4,and IL-10 were also evaluated.

Results

Immunization with rEg.P29 induced protective immune responses up to 94.5 %, compared with immunoadjuvant group. The levels of specific IgG, IgG1, IgG2, and IgE as well as IFN-γ, IL-2, and IL-4 significantly increased after two immunizations (P < 0.05); however, the levels of IgM and IL-10 did not show difference.

Conclusion

rEg.P29 showed Immunoprotection and induced Th1 and Th2 immune responses; hence, rEg.P29 is a potential vaccine for E. granulosus infection.

     

Antimicrobial susceptibility and phylotyping profile of pathogenic <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Salmonella enterica</Emphasis> isolates from calves and pigs in Minas Gerais,Brazil

The aims of the present study were to determine (i) the profiles of phylogroup and (ii) the antimicrobial susceptibility of pathogenic Escherichia coli strains isolated from calves, and of Salmonella spp. strains isolated from calves and pigs in Minas Gerais State, Brazil. Sixty-one pathogenic E. coli strains and Salmonella spp. (n?=?24) strains isolated from fecal samples of calves and Salmonella spp. (n?=?39) strains previously isolated from fecal samples of growing/finishing pigs were tested. The minimum inhibitory concentration (MIC) using the agar dilution method was determined for nalidixic acid, amikacin, amoxicillin, ampicillin, cefoxitin, norfloxacin, gentamicin, tetracycline, and trimethoprim-sulfamethoxazole. All E. coli isolates were susceptible to amikacin. Tetracycline was the antimicrobial that presented the higher frequency of resistance among E. coli strains, followed by ampicillin, trimethoprim-sulfamethoxazole, amoxicillin, nalidixic acid, norfloxacin, gentamicin, and cefoxitin. E. coli (n?=?61) strains isolated from calves belonged to different phylogroup namely, phylogroup A (n?=?26), phylogroup B1 (n?=?31), phylogroup E (n?=?3), and phylogroup F (n?=?1). Phylogroups B2, C, and D were not identified among the E. coli in the present study. All Salmonella spp. (n?=?24) strains isolated from fecal samples of calves were susceptible to amikacin, amoxicillin, ampicillin, norfloxacin, gentamicin, tetracycline, and trimethoprim-sulfamethoxazole. Resistance to nalidixic acid and cefoxitin was detected in 16.66 and 8.33 % of the Salmonella spp. strains, respectively. Among the Salmonella spp. (n?=?39) strains isolated from fecal samples of pigs, the higher frequency of resistance was observed to tetracycline, followed by amoxicillin, gentamicin, ampicillin, trimethoprim-sulfamethoxazole, nalidixic acid, cefoxitin, and norfloxacin. All strains were susceptible to amikacin. Forty-eight (78.68 %) of the E. coli strains were classified as multidrug-resistant, whereas among Salmonella spp. strains, the percentage of multidrug resistance was 57.14 %, being all multidrug-resistant strains isolated from pigs (92.30 %). The results from the present study indicate a high frequency of antimicrobial resistance among pathogenic E. coli strains isolated from calves and Salmonella spp. strains isolated from pigs and a high rate of susceptibility to most antimicrobials tested among Salmonella spp. strains isolated from calves. Our study highlights the presence of multidrug-resistant strains of E. coli and Salmonella spp. isolated from food-producing animals in Minas Gerais, Brazil.     

Microbial community and ovine host response varies with early and late stages of <Emphasis Type="Italic">Haemonchus contortus</Emphasis> infection

The interactions between gastric microbiota, ovine host, and Haemonchus contortus portray the ovine gastric environment as a complex ecosystem, where all factors play a pertinent role in fine-tuning each other and in haemeostasis. We delineated the impact of early and late Haemonchus infection on abomasal and ruminal microbial community, as well as the ovine host. Twelve, parasite-naive lambs were divided into four groups, 7 days post-infection (dpi) and time-matched uninfected-control groups; 50 dpi and time-matched uninfected control groups were used for the experiment. Six sheep were inoculated with 5000 H. contortus infective larvae and followed for 7 or 50 days with their corresponding uninfected-control ones. Ovine abomasal tissues were collected for histological analysis and gastric fluids were collected for PH value measurements, microbial community isolation and Illumina MiSeq platform and bioinformatic analysis. Our results showed that Haemonchus infection increased the abomasal gastric pH (P = 0.05) and resulted in necrotizing and inflammatory changes that were more severe during acute infection. Furthermore, infection increased the abomasal bacterial load and decreased the ruminal microbiome. A 7-day infection of sheep with H. contortus significantly altered approximately 98% and 94% of genera in the abomasal and ruminal bacterial profile, respectively (P = 0.04–0.05). However, the approximate altered genera 50 days after infection in the ovine abomasal and ruminal microbiome were about 62% and 69%, correspondingly (P = 0.04–0.05) with increase in some bacteria and decrease in others. Overall, these results indicate that Haemonchus infection plays a crucial role in shaping stomach microbial community composition, and diversity.     

Performance and carcass characteristics of lambs fed diets with increasing levels of <Emphasis Type="Italic">Mimosa tenuiflora</Emphasis> (Willd.) hay replacing Buffel grass hay

This study evaluated the performance and carcass characteristics of lambs fed diets with increasing levels of Mimosa tenuiflora (Willd.) hay replacing Buffel grass (Cenchrus ciliaris). Twenty-eight Santa Inês male lambs with an average body weight (BW) of 20.3 ± 1.49 kg(mean ± SD) were allocated in individual stalls and distributed in a completely random design with four treatments (0, 20, 40, and 60 g/100 g total DM M. tenuiflora hay replacing Buffel grass hay in diet) with seven replications. M. tenuiflora hay at the level of 20% dry matter (DM) total replacing Buffel grass hay increased final weight (P = 0.006), total weight gain (P < 0.001), average daily weight gain (ADWG; P < 0.001), DM intake (P < 0.001), and feed efficiency (P < 0.001). Intake of crude protein, NDF_ap, ADF_ap, ash, ether extract, total and non-fibrous carbohydrates, and total digestible nutrients presented a positive quadratic effect with M. tenuiflora hay replacing Buffel grass hay and 40 g/100 g total DM level presented greater intake. There were positive quadratic effects by M. tenuiflora hay inclusion at 20 g/100 g total DM level on slaughtering weight (P = 0.005), hot carcass weight (P = 0.002), cold carcass weight (P = 0.002), empty body weight (P = 0.001), hot carcass yield (P = 0.002), cold carcass yield (P = 0.003), and increase linear on biological yield (P = 0.003). There was no influence on cooling weight loss (P = 0.284). M. tenuiflora hay may be included in lamb diets at amounts up to 20 g/100 g total DM substitution of Buffel grass hay because increase in the nutrients intake, growth performance, and carcass characteristics.     

Effect of grape seed and skin supplement on milk yield and composition of dairy ewes

In the present study, we investigated the effect of grape seed and skin supplement (GSSS), on lactating dairy ewes’ production. Ten dairy pregnant ewes from northern Tunisia were allocated to two groups: control diet (C) and supplemented with 20 % (w/w) GSSS. The experiment lasted for 8 weeks and took place after 2 months of lambing. During the experiment, daily milk yield and milk composition were determined. Supplementation of the diet with GSSS increased milk production (P?<?0.001), calcium (P?<?0.01), free iron (P?<?0.01) and urea content (P?<?0.001) but had no effect on milk fat nor protein. From these data, it is concluded that the inclusion of GSSS in sheep diets increased significantly ewes’ milk yield.     

Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Comparison of milk fatty acid profiles measured on Kouri cows near Lake Chad and on dairy cattle as reported by meta-analytical data

Kouri (Bos taurus) is a breed aboriginal from Lake Chad and threatened with extinction. This study aimed to compare milk fatty acid profiles measured on Kouri cows and on high-yielding dairy cattle in Europe and elsewhere as reported by meta-analytical data (22 experimentations). Milk samples were collected from 14 Kouri dairy cows in dry season (March to June) and fatty acids (FA) were determined by gas chromatography. Overall, 32 FA have been identified. Kouri showed lower values (P < 0.001) in the sum of saturated FA (SFA, ?10.9 pts), cis-9, cis-12 18:2 (?1.00 pt) (P < 0.01, higher values (P < 0.001) in the sum of monounsaturated FA (MUFA, +15.3 pts), C18:0) (+3.5 pts), cis-9, trans-11 C18:2-CLA (+1.00 pts), trans-11 18:1 (+1.4 pts) and (P < 0.01) in cis-9, C18:1 (+3.00 pts) acids. The differences between the milk FA profile of the Kouri cows and that obtained from meta-analytical data could be the possible consequence of the use of particular lake pastures by Kouri cows.     

<Emphasis Type="SmallCaps">d</Emphasis>-Cloprostenol enhances estrus synchronization in tropical hair sheep

To compare the effects of PGF2α (dinoprost tromethamine) and d-cloprostenol in a two-dose protocol for estrus synchronization in hair sheep during breeding season in Yucatán, México, two experiments were conducted. In experiment 1, 61 cyclic hair sheep were divided into two groups: G1 (control n?=?30), two doses of 50 μg of dinoprost tromethamine IM with 12 days between applications, and G2 (n?=?31), two doses of 50 μg of d-cloprostenol IM at the same time interval. For determination of progesterone levels, 16 ewes from each group were randomly selected. In experiment 2, 70 cyclic hair sheep were assigned at the same treatments (G1 and G2, n?=?35) and 48 h after the second application, the ewes in estrus were detected by two vasectomized rams. Sheep with detected estrus were inseminated, and 45 days after, pregnant animals were identified by ultrasonography. An exact Fisher’s test was performed for the analysis of ewes in estrus (experiments 1 and 2) and number of pregnant ewes (experiment 2); for the comparison of time between end of treatment-estrus presentation, a survival analysis was used. Duration of estrus in hours was analyzed using a generalized mixed model (GLM) ANOVA whereas plasma progesterone concentrations were analyzed by non-linear regression. There were significant differences (P?<?0.05) in the proportion of ewes in estrus upon treatments (G1, 57% vs G2, 87% and G1, 37.1% vs G2, 65.7% in experiments 1 and 2, respectively), and between the end of treatment-onset estrus interval (P?<?0.01), survival curves showed the highest number of sheep in estrus between 40 and 48 h (G1, 43.7?+?8.05 h vs G2, 42.9?+?6.7 h, experiment 1). There were no significant differences (P?>?0.05) in duration of estrus (G1, 42?+?6.1 h, vs G2, 41.1?+?11.2 h, experiment 1) and pregnancy in the ewes that presented estrus, and were inseminated (G1, 38.4% vs 52.1%, experiment 2). With regard to concentrations of progesterone, significant differences (P?<?0.01) were found between treatments, and progesterone levels before the second application of d-cloprostenol were higher. In consideration of the results, it can be concluded that in a two-dose protocol of a luteolytic agent, more ewes presented estrus in response to d-cloprostenol compared to dinoprost tromethamine with similar pregnancy rates.     

Oocyte maturation,embryo development and gene expression following two different methods of bovine cumulus-oocyte complexes vitrification

Objective

To examine the maturational competence, embryo development and expression of genes involved in oocyte maturation and cumulus expansion (GDF9, BMP15, HAS2, TNFAIP6, FGF17 and FSHr) following two standard methods of bovine COCs vitrification.

Methods

Bovine cumulus-oocyte complexes (COCs) were aspirated from slaughtered ovaries and then distributed into three groups: non-vitrified COCs (control), vitrification 1 group (V1); vitrification was performed by 15% ethylene glycol (EG) and 15% DMSO in holding media (TCM-199 with 20% FCS); and vitrification 2 group (V2); vitrification was performed by 40% EG in holding media. After vitrification, COCs were warmed in two steps and cultured and then evaluated for nuclear maturation, embryo development and gene expressions.

Results

The mean (±SD) percentages of nuclear maturation and blastocyst/cleaved were higher in control group (79.5 ± 8.0 and 31.0 ± 5.1%) than the V1 (34.8 ± 9.1 and 4.4 ± 5.1%) and V2 (47.8 ± 11.7 and 7.1 ± 5.8%) groups (P < 0.05), respectively. Further, COCs in V2 group showed higher mean (±SD) percentages of cleavage compared to V1 group (31.8 ± 1.0 vs 21.7 ± 2.8%; P < 0.05). GDF9 and BMP15 expression levels were higher in COCs in the control than of the vitrification groups (P < 0.05). In addition, expression level of GDF9 and BMP15 was higher in V2 group than in V1group (P < 0.05). The expression of HAS2 and FGF17 in V1 group was lower (P < 0.05) than that of the V2 groups.

Conclusions

Expression of oocyte maturation genes was affected by vitrification procedure and conditions. Using EG alone for vitrification of bovine immature COCs, resulted in higher expression of GDF9, BMP15 and production of more in vitro matured and cleaved oocytes.