<Emphasis Type="Italic">Arcobacter</Emphasis> spp. in fecal samples from Brazilian farmed caimans (<Emphasis Type="Italic">Caiman yacare</Emphasis>, Daudin 1802)

The aim of this study was to perform the identification and molecular characterization of Arcobacter cryaerophilus and Arcobacter butzleri isolated from caiman (Caiman yacare), kept at a production farm, in Brazil. Forty fecal samples were analyzed. After isolation and identification, 21/40 strains of A. butzleri and 19/40 strains of A. cryaerophilus were subjected to PCR for potential virulence gene detection. The results of the PCR showed 38/40 strains positive for the cadF, cj1349, ciaB, and tlyA genes, 39/40 strains positive for the pldA gene, and 40/40 strains positive for the mviN gene. None of the strains presented the irgA gene. Hemagglutinin (hecA gene) and hemolysin (hecB) genes were detected in 21/40 and 16/40 strains, respectively. The SE-AFLP showed a great genetic diversity, but some clonally groups were disseminated in various tanks. These data reveal that the strains presented the same virulence traits described from Arcobacter isolated from food-borne disease in humans.     

Evaluation of PCR methods for detection of <Emphasis Type="Italic">Brucella</Emphasis> strains from culture and tissues

The genus Brucella causes significant economic losses due to infertility, abortion, stillbirth or weak calves, and neonatal mortality in livestock. Brucellosis is still a zoonosis of public health importance worldwide. The study was aimed to optimize and evaluate PCR assays used for the diagnosis of Brucella infections. For this aim, several primers and PCR protocols were performed and compared with Brucella cultures and biological material inoculated with Brucella. In PCR assays, genus- or species-specific oligonucleotide primers derived from 16S rRNA sequences (F4/R2, Ba148/928, IS711, BruP6-P7) and OMPs (JPF/JPR, 31ter/sd) of Brucella were used. All primers except for BruP6-P7 detected the DNA from reference Brucella strains and field isolates. In spiked blood, milk, and semen samples, F4-R2 primer-oriented PCR assays detected minimal numbers of Brucella. In spiked serum and fetal stomach content, Ba148/928 primer-oriented PCR assays detected minimal numbers of Brucella. Field samples collected from sheep and cattle were examined by bacteriological methods and optimized PCR assays. Overall, sensitivity of PCR assays was found superior to conventional bacteriological isolation. Brucella DNA was detected in 35.1, 1.1, 24.8, 5.0, and 8.0% of aborted fetus, blood, milk, semen, and serum samples by PCR assays, respectively. In conclusion, PCR assay in optimized conditions was found to be valuable in sensitive and specific detection of Brucella infections of animals.     

Comparison of rapid diagnostic tests to detect <Emphasis Type="Italic">Mycobacterium avium</Emphasis> subsp. <Emphasis Type="Italic">paratuberculosis</Emphasis> disseminated infection in bovine liver

Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s disease, a chronic enteritis in cattle and other domestic and wild ruminants. The presence of MAP in tissues other than intestines and associated lymph nodes, such as meat and liver, is a potential public health concern. In the present study, the relationship between the results of rapid diagnostic tests of the Johne’s disease, such as serum ELISA, rectal scraping PCR, and acid-fast staining, and the presence of MAP in liver was evaluated. Blood, liver, and rectal scraping samples were collected from 200 slaughtered cattle with unknown Johne’s disease status. ELISA was performed to determine the MAP antibody activity in the serum. Acid-fast staining was performed on rectal scraping samples, and PCR was performed on rectal scraping and liver samples. PCR-positive liver samples were used for mycobacterial culture. Overall, the results of this study demonstrated that MAP can be detected and cultured from liver of slaughtered cattle and rapid diagnostic tests of Johne’s disease have limited value in detecting cattle with MAP infection in liver. These findings show that the presence of MAP in liver tissue may occur in cows with negative results for rapid diagnostic tests and vice versa. Hence, liver might represent another possible risk of human exposure to MAP. Given concerns about a potential zoonotic role for MAP, these results show the necessity to find new methods for detecting cattle with MAP disseminated infection.     

Prevalence and risk factors associated with <Emphasis Type="Italic">Theileria parva</Emphasis> infection in cattle in three regions of Tanzania

Ticks and tickborne diseases (TBDs) are serious constraints to cattle production in Tanzania and other tropical and subtropical countries. Among the TBDs, East Coast fever (ECF) is the most important as it causes significant economic losses to the cattle industry in Tanzania. However, control of ECF in Tanzania has continued to be a challenge due to inadequate epidemiological information. The main objective of this study was to determine the epidemiological situation of Theileria parva infections in cattle kept under pastoral and agro-pastoral farming systems in Mara, Singida, and Mbeya regions of Tanzania. Blood samples were collected from 648 cattle in the three regions. Genomic DNA was extracted and amplified in a polymerase chain reaction (PCR) using T. parva-specific primers targeting the 104-kD antigen (P104) gene. In addition, information was collected on the possible risk factors of T. parva infection (animal age, region, animal sex, tick burden, tick control method, and frequency of acaricide application). The prevalence of T. parva across the three regions was 14.2%. There was variation in prevalence among the three regions with Mara (21.8%) having a significantly higher (p = 0.001) prevalence than the other regions. Moreover, Mbeya exhibited relatively lower prevalence (7.4%) compared to the other regions. Factors found to be significantly associated with an animal being PCR positive for T. parva were region (p = 0.001) and tick burden (p = 0.003). Other factors were not found to be significant predictors of being PCR positive for T. parva. The present study showed high variation in tick burden and T. parva prevalence across the regions. Therefore, different strategic planning and cost-effective control measures for ticks and T. parva infection should be implemented region by region in order to reduce losses caused by ticks and ECF in the study area.     

Detection of <Emphasis Type="Italic">Trypanosoma vivax</Emphasis> DNA in semen from experimentally infected goats

The present work aimed to investigate the presence of T. vivax DNA in the semen of experimentally infected goats. Twelve male goats native to the Brazilian Northeast, adults, were randomly assigned to two experimental groups: the infected group consisting of six goats infected intravenously with 0.5 mL of blood containing approximately 1.25?×?10⁵ trypomastigotes of T. vivax, and a control group composed of six uninfected goats. After the infection, clinical examinations aiming to evaluate rectal temperature, parasitemia and hematocrit were performed. Semen samples were collected from goats by electroejaculation on the 7th, 14th and 21st days post-infection (dpi). The recombinant DNA-encoding gene encoding the L-like-specific gene for T. vivax. The infection was characterized by increased rectal temperature, high parasitemia and significant reduction of hematocrit values. Results for T. vivax DNA detection using TviCatL-PCR were positive in all semen samples from the infected group collected on 7th, 14th and 21st dpi. The presence of T. vivax DNA in 7th dpi suggests the early invasion of the parasite in the reproductive organs. Also, the finding of T. vivax DNA in all periods analyzed may suggest the continued elimination of the parasite in the semen, which may increase the chances of sexual transmission. Thus, T. vivax DNA is recorded for the first time in the semen of infected goats. Thus, these data are of great importance, since the detection of the T. vivax genetic material in the semen may point to the possibility that the parasite may be transmitted through the sexual pathway.     

Influence of <Emphasis Type="Italic">Nigella sativa</Emphasis> seeds, <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis> leaves and their combination on growth performance,immune response and rumen metabolism in Dorper lambs

The objective of this study was to determine the effects of dietary supplementation of Nigella sativa L. seeds, Rosmarinus officinalis L. leaves and their combination on rumen metabolism, nutrient intake and digestibility, growth performance, immune response and blood metabolites in Dorper lambs. Twenty-four entire male Dorper lambs (18.68?±?0.6 kg, 4–5 months old) were randomly assigned to a concentrate mixture containing on a dry matter basis either, no supplement (control, T1), 1% R. officinalis leaves (T2), 1% N. sativa seeds (T3) or 1% R. officinalis leaves +1% N. sativa seeds (T4). The lambs had ad libitum access to urea-treated rice straw (UTRS) and were raised for 90 days. Supplemented lambs had greater (P?< 0.05) intake of DM and UTRS than the control lambs. The T4 lambs had lower (P?< 0.05) nutrient digestibility than those fed other treatments. Total and daily weight gain was greater (P?< 0.05) in T2 lambs than those fed other diets. The T3 and T4 lambs had greater (P?< 0.05) ruminal pH than the T1 and T2 lambs. Supplemented lambs had lower (P?<?0.05) ruminal total volatile fatty acids, acetate, propionate, NH₃-N and C18:0 than the control lambs. The T4 lambs had lower (P?< 0.05) population of Fibrobacter succinogenes, Ruminococcus albus, methanogens and total protozoa compared with those fed other diets. Supplemented lambs had lower (P?< 0.05) neutrophils, basophils and serum urea and greater (P?<?0.05) serum IgA and IgG compared with the control lambs. The current results emphasised the variation in the efficacy of medicinal plants in ruminant nutrition.     

Sample pooling for real-time PCR detection and virulence determination of the footrot pathogen <Emphasis Type="Italic">Dichelobacter nodosus</Emphasis>

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.     

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.     

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.     

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.

     

Seroprevalence of <Emphasis Type="Italic">Anaplasma marginale</Emphasis> and <Emphasis Type="Italic">Babesia bigemina</Emphasis> infections and associated risk factors in Machakos County,Kenya

Anaplasma marginale and Babesia bigemina are important tick-borne pathogens of cattle. A cross-sectional survey was undertaken to determine the seroprevalence of A. marginale and B. bigemina infections and identify associated risk factors on traditional smallholder farms in Machakos County, Kenya. A total of 421 cattle from 127 farms from four divisions in the county were sampled and visited between September and November 2007. The farms were selected by a proportional allocation approach based on the number of farms in the four divisions previously selected by stratified random sampling method. Information on animal and individual farm management variables was obtained using standardized questionnaires. Prevalence of serum antibodies due to A. marginale and B. bigemina pathogens was determined using the enzyme-linked immunosorbent assay (ELISA) technique. The relationship between the seropositivity and associated risk factors was assessed by multivariable analyses using standard logistic regression models. The overall estimation (and their 95% confidence intervals) of A. marginale and B. bigemina seropositivity at the animal level was 53.4% (48.5%, 58.2%) and 40.6% (35.8%, 45.4%), respectively. Two variables, “animal age” and “administrative division,” were significantly associated with the A. marginale seroresponse. Three variables, “animal age” “grazing system” and “administrative division” were significantly associated with the B. bigemina seroresponse. These findings suggest possible indicators of existence of endemic instability for the two infections. The study identifies characterization of environmental suitability for the vectors and how they interact with grazing systems to cause the infections as an area for further studies, for improved understanding of the infections and in designing disease control programs.     

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.     

Control of <Emphasis Type="Italic">Haemonchus contortus</Emphasis> in sheep using basidiocarps of <Emphasis Type="Italic">Agaricus blazei</Emphasis> Murril

Objective

This study evaluated the effects in vitro and in vivo of Agaricus blazei against Haemonchus contortus in sheep.

Methods

The in vitro efficacy of aqueous extract on egg hatching inhibition (EHI) was investigated and after 72 h incubation with varying concentrations the effects on, blastomeres, embryonated eggs, and first stage larvae (L1) were evaluated. Larval development inhibition (LDI) for dry powder and the aqueous extract were evaluated in fecal cultures of sheep infected with H. contortus. In vivo efficacy was determined by reduction in fecal egg count (FEC). Lambs were treated with powder A. blazei (11.4 g/kg pc) or trichlorfon, or were untreated and the possible toxicity of this fungus was monitored by plasmatic enzyme analysis.

Results

Concentrations equal to and higher than 3.62 mg/mL and of aqueous extract were 100% effective in the EHI test. In the LDI test, LC90 was estimated for 5.66 and 106.0 mg/g fecal culture for aqueous extract and powder, respectively. The mean FEC in lambs 14 days post-treatment with A. blazei powder was significantly lower than observed for the negative control, and the serum levels of aspartate transaminase and alanine transaminase were normal.

Conclusion

The fungi supplementation promotes, respectively, high and moderate anthelmintic efficacy with in vitro and in vivo tests, respectively, suggesting it as an alternative or complementary treatment for haemonchosis in sheep.

     

Multidrug-resistant <Emphasis Type="Italic">Salmonellae</Emphasis> isolated in Japanese quails reared in Abeokuta,Nigeria

Salmonellosis is a major bacterial disease causing huge economic losses in the poultry industry worldwide. This study was carried out to determine the period prevalence and antimicrobial susceptibility of Salmonella enterica in Japanese quails in Abeokuta, Nigeria. Four hundred cloacal swabs of quail birds were collected from 4 locations within Abeokuta. Salmonella was isolated from the samples using conventional methods for selective isolation of Salmonella and biochemical identification. Isolates were confirmed by polymerase chain reaction assays for the amplification and detection of Salmonella-associated virulence genes (invA and stn) using specific primers. Antimicrobial susceptibility testing was done using the Kirby-Bauer disk diffusion method. In all, Salmonella was isolated from 14 (3.5%) cloacal swabs. All 14 isolates possessed invA and stn genes. The Salmonella isolates showed resistance to tetracycline (100%), doxycycline (100%), ampicillin (100%), sulphamethoxazole (92.9%), nalidixic acid (85.8%), ceftazidime (78.6%), neomycin (64.3%), streptomycin (50%) and gentamycin (28.6%) but all the isolates were susceptible to ciprofloxacin. The isolates were resistant to at least three antimicrobials indicating multidrug resistance. The results concluded that Japanese quails harbour multidrug-resistant Salmonella which could be transmitted to humans through consumption of contaminated food or by direct and indirect contact with the carrier birds. Antimicrobial resistance could be due to overdependence on antimicrobials. Ciprofloxacin could be considered in the treatment of zoonotic Salmonellosis in humans.     

Virulence traits of avian pathogenic (APEC) and fecal (AFEC) <Emphasis Type="Italic">E. coli</Emphasis> isolated from broiler chickens in Algeria

Avian pathogenic E. coli (APEC) is the etiologic agent of avian colibacillosis, the most common disease responsible for chicken morbidity in the world. Although multiple virulence-associated factors were identified, their prevalence in Algeria is still poorly known. In the present research, 92 avian pathogenic E. coli (APEC) isolates were recovered from broilers with clinical signs and lesions of colibacillosis. In addition, 32 E. coli isolates collected from feces of healthy birds (AFEC) were included for comparison. All isolates were investigated by PCR for the presence of a total of 11 virulence-associated genes described for avian pathogenic (iroN, ompT, hlyF, iss, iutA, and fimC) and diarrheagenic E. coli (eae, stx, elt/est, ipaH, and aggR). The sensitivity of 39 APEC isolates to 16 antibiotics was also determined using antimicrobial pretreated microplates. Here, we report that 98% of the examined isolates host at least one of the tested virulence factors. The most prevalent genes in APEC were iutA (90.6%), ompT (86.9%), and iss (85.8%); whereas, iutA (78.1%), fimC (78.1%), and iroN (68.7%) were the highest prevalent genes in AFEC. Our data showed that none of the AFEC isolates harbor any of the tested diarrheagenic genes. Moreover, only elt/est (5.4%), stx (2.1%), and ipaH (2.1%) genes were carried by APEC isolates. We further established that ceftazodime, ceftiofur, mequindox, amoxicillin/clavulanic acid, and meropenem were the most efficient antibiotics against the analyzed APEC isolates. Overall, our findings provide more insights about APEC and AFEC virulence potential in Algeria which could participate in the fight against colibacillosis.     

Effect of feed restriction on intake of <Emphasis Type="Italic">Moringa oleifera</Emphasis> and <Emphasis Type="Italic">Leucaena leucocephala</Emphasis> and growth performance of rabbits

Two experiments were conducted to evaluate the effect of feed restriction on intake of Moringa oleifera (MO) or Leucaena leucocephala (LL) and growth of rabbits. In experiment one, 45 rabbits (male and female) weighing 1.18 ± 0.15 kg were used. They were randomly distributed to three feed restriction treatments (20, 30, and 40%) with 15 rabbits each (9 females and 6 males) and they were offered M. oleifera (MO) ad libitum. In experiment two, 45 growing male rabbits weighing 0.63 ± 0.113 kg were used. They were randomly assigned to 0, 20, and 30% feed restriction diets, and they have free access to L. leucocephala (LL). Intake of MO increased (P < 0.05) conforming feed restriction increased (40.6, 52.9, and 55.2 g/day of MO for 20, 30, and 40%, respectively). Daily liveweight gain and feed conversion did not differ (P > 0.05), and economic efficiency was similar among treatments. Consumption of LL increased (P < 0.05) in rabbits under the 30% restriction treatment in comparison to that of rabbits restricted 20% (46.0 and 44.4 g/day, respectively). Total feed intake (LL + feed) was highest in 20% restricted rabbits (108.0, 100.8, and 93.2 g/day for 20, 30, and 0%, respectively). Daily liveweight gain and feed conversion were not affected by feed restriction (P > 0.05). Economic efficiency improved twice in feed-restricted rabbits (2.0 and 2.3 for 20 and 30%, respectively) in contrast to that of the control 0% group (1.1). The results suggest that rabbits restricted up to 30% and supplemented with either MO or LL did not affect growth performance and reduced feed cost.     

Efficacy of eprinomectin against <Emphasis Type="Italic">Toxocara vitulorum</Emphasis> in calves

This study was made to investigate efficacy of eprinomectin pour-on against to Toxocara vitulorum in calves. In the study, 16 calves naturally infected with T. vitulorum were divided into two groups as treatment (eight calves) and control (eight calves). Eprinomectin (0.5 mg/kg, Eprinex®, Merial) was given to treatment group calves, and eggs per gramme were determined in the faeces on the day of pre-treatment and the second, third, fourth, fifth, sixth, 14th and 28th days of post-treatment. No side effects associated with nervous, respiratory and gastrointestinal systems were observed. In conclusion, eprinomectin was determined to be 100% effective against T. vitulorum. This is the first study to evaluate the efficacy of eprinomectin against a natural T. vitulorum infection in calves.     

<Emphasis Type="Italic">In vitro</Emphasis> larvicidal effect of a hydroalcoholic extract from <Emphasis Type="Italic">Acacia cochliacantha</Emphasis> leaf against ruminant parasitic nematodes

The aim of this study was to evaluate the in vitro lethal effect of a hydroalcoholic extract (HAE) from Acacia cochliacantha leaf against three gastrointestinal nematodes species (Haemonchus contortus, H. placei and Cooperia punctata) of domestic ruminants. The HAE was assessed using five concentrations: 100, 125, 175, 150 and 200 mg/ml; 0.5% Ivermectin was used as a positive control and distilled water, as negative control. The data were normalized using the square root and analysed with a completely randomized design through ANOVA analysis using the general lineal model (GLM) of the SAS program. The HAE tannin content was determined through spectrophotometry (UV-visible) and the other major phenols, were identified by chromatographic processes. The results showed an in vitro larvicidal activity of the HAE against the three assessed nematode species with all assessed concentrations. A clear HAE increased concentration dependence effect was observed. The highest activity of the HAE was obtained at the highest concentration (close to 100%, P < 0.05). This result was similar to the one obtained with Ivermectin. On the other hand, the chemical analysis of HAE showed the presence of tannins, caffeoyls and coumaroyl derivates and quercetin as the main compounds. The results suggest that the HAE from this plant species possess in vitro anthelmintic properties. The identified compounds in this study would good candidates for further in vivo researches.     

Genotypic analysis of virulence genes and antimicrobial profile of diarrheagenic <Emphasis Type="Italic">Escherichia coli</Emphasis> isolated from diseased lambs in Iran

The aim of the present study was to determine the analysis of virulence genes and antimicrobial profile of diarrheagenic Escherichia coli isolated from diseased lambs. Two hundred ninety E. coli isolates were recovered from 300 rectal swabs of diarrheic lambs and were confirmed by biochemical tests. The pathotype determination was done according to the presence of genes including f5, f41, LTI, STI, bfp, ipaH, stx ₁ , stx ₂ , eae, ehlyA, cnf ₁ , cnf ₂ , cdIII, cdIV, and f17 by PCR method. Sixty-six isolates (23.72%) possessed the STI gene and categorized into entrotoxigenic E. coli (ETEC). Nine isolates (3.1%) and five isolates (1.72%) were positive for the cnf1 and cnf2 genes which categorized into necrotoxic E. coli (NTEC). Hundred and seventeen isolates (40.34%) harbored stx ₁ and/or stx ₂ and classified as Shiga toxin-producing E. coli (STEC). Thirteen isolates (4.48%) were assigned to atypical entropathogenic E. coli (aEPEC) and possessed eae gene. Two isolates (0.68%) were positive for ipaH gene and were assigned to entroinvasive E. coli (EIEC). Statistical analysis showed a specific association between eae gene and STEC pathotype (P?<?0.0001). The most prevalent resistance was observed against lincomycin (96.5%) and the lowest resistance was against kanamycine (56.02%), respectively. The high prevalence of STEC and ETEC indicates that diarrheic lambs represent an important reservoir for humans. ETEC may play an important role for frequent occurrence of diarrhea in lambs observed in this region. Due to high antibiotic resistance, appropriate control should be implemented in veterinary medicine to curb the development of novel resistant isolates.