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.     

Characterization of non-O157 shiga toxin-producing Escherichia coli isolates from healthy fat-tailed sheep in southeastern of Iran

The objectives of this study were to determine the presence and prevalence of non-O157 shiga toxin-producing Escherichia coli (STEC) isolates from faeces of healthy fat-tailed sheep and detection of phylogenetic background and antibiotic resistance profile of isolates. One hundred ninety-two E. coli isolates were recovered from obtained rectal swabs and were confirmed by biochemical tests. Antibiotic resistance profiles of isolates were detected and phylogenetic background of isolates was determined according to the presence of the chuA, yjaA and TspE4.C2 genetic markers. The isolates were examined to determine stx ₁ , stx ₂ and eae genes. Non-O157 STEC isolates were identified by using O157 specific antiserum. Forty-three isolates (22.40 %) were positive for one of the stx ₁ , stx ₂ and eae genes, whereas 10.42 % were positive for stx ₁ , 19.38 % for eae and 2.60 % for stx ₂ gene. None of the positive isolates belonged to O157 serogroup. Twenty isolates possessed stx ₁ were distributed in A (six isolates), B1 (13) and D (one) phylogroups, whereas stx ₂ positive isolates fell into A (three isolates) and B1 (two) phylogenetic groups. Eighteen isolates contained eae gene belonged to A (five isolates), B1 (seven) and D (six) phylogroups. The maximum and minimum resistance rates were recorded against to penicillin and co-trimoxazole respectively. The positive isolates for stx ₁ , stx ₂ and eae genes showed several antibiotic resistance patterns, whereas belonged to A, B1 and D phylogroups. In conclusion, faeces of healthy sheep could be considered as the important sources of non-O157 STEC and also multidrug-resistant E. coli isolates.     

Virulence repertoire of Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic Escherichia coli (ETEC) from diarrhoeic lambs of Arunachal Pradesh,India

A total of 107 faecal samples were collected from diarrhoeic lambs of high altitude terrains (2,000 to 5,000 m above the mean sea level) of Tawang and West Kameng districts of Arunachal Pradesh, India. Total 234 Escherichia coli were isolated and further subjected to PCR for the study of virulence repertoire characteristics of Shiga toxin-producing E. coli (STEC) and enterotoxigenic E. coli (ETEC). Out of the 234 isolated E. coli, 32% were found positive for STEC, and 9% were carrying virulence gene for ETEC. The isolated STEC serogroups were O159, O127, O120, O113, O60, O30, O25, O8 and O2. Of all the 74 STEC strains, PCR showed that 18% isolates carried stx ₁ , 26% possessed stx ₂ and 47% produced positive amplicon for both. Other virulent attributes like intimin (eaeA), enterohaemolysin (ehxA) and STEC auto-agglutinating adhesin (saa) were present in 18%, 43% and 44% of the isolates, respectively. The isolated ETEC serogroups were O172, O170, O159, O146, O127, O120, O113, O86, O75, O60, O30, O25, O8, O2, OR and OUT. Of the 22 ETEC-positive isolates, 23%, 18% and 4.5% possessed the gene only for LT, STa and STb, respectively, whereas 54% carried genes for both LT and STb. Some serogroups of E. coli like O159, O127, O120, O113, O60, O30, O25, O8 and O2 possessed genes for both Shiga toxin and enterotoxin. This study is the first report of ETEC isolation from diarrhoeic lambs in India. The moderately high proportion of STEC and ETEC in the diarrhoeic lambs implicated that these animals are important reservoir of STEC and ETEC. This is really a grave concern for the ‘brokpas’ and nomads (shepherds) who share a close relationship with this animals for their livelihood. This study also indicates that ETEC may be a major cause for frequent diarrhoeal episodes in lambs of this region.     

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.     

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.     

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.     

Isolation,identification and differentiation of <Emphasis Type="Italic">Campylobacter</Emphasis> spp. using multiplex PCR assay from goats in Khartoum State,Sudan

The aim of this study was to identify and characterize thermophilic Campylobacter species in faecal samples from goats in Khartoum State, Sudan, by application of multiplex polymerase chain reaction. Campylobacteriosis is a zoonotic disease of global concern, and the organisms can be transmitted to human via food, water and through contact with farm animals and pets. There are five clinically related Campylobacter species: Campylobacter jejuni (C. jejuni). Campylobacter coli, Campylobacter lari, Campylobacter upsaliensis and Campylobacter fetus. Conventional cultural methods to diagnose campylobacteriosis are tedious and time consuming. Wide ranges of genes have been reported to be used for PCR-based identification of Campylobacter spp. We used a multiplex PCR assay to simultaneously detect genes from the major five clinically significant Campylobacter spp. The genes selected were hipO (hippuricase) and 23S rRNA from glyA (serine hydroxymethyl transferase) from each of C. jejuni. C. coli, C. lari, and C. upsaliensis; and sapB2 (surface layer protein) from C. fetus subsp. fetus. The assay was used to identify Campylobacter isolates recovered from 336 cultured faecal samples from goats in three localities in Khartoum State. C. coli was the most predominant isolate (234; 69.6%), followed by C. jejuni (19; 5.7%), C. upsaliensis (13; 3.9%), C. fetus subsp. fetus (7; 2.1%) and C. lari (6; 1.8%). Twenty-nine goats showed mixed infection with Campylobacter spp., 21 of which harbored two Campylobacter spp., while eight animals were infected with three species. Ten out of twelve goats that displayed diarrhea harbored C. coli only. C. coli, C. jejuni and C. upsaliensis showed significant variation with localities. The prevalence of C. coli was significantly higher (87; 25.9%) in goats from Omdurman, whereas C. jejuni and C. upsaliensis were significantly higher (11; 3.3%, 9; 2.7%) in goats from Khartoum. The multiplex PCR assay was found to be rapid and easy to perform and had a high sensitivity and specificity for characterizing the isolates, even in mixed cultures. The study demonstrated the significance of goats as reservoirs in the dissemination of Campylobacter spp. which could be considered as potential agent of caprine enteritis and abortion as well as contamination of the wider environment posing serious public health concern in Khartoum State.     

Hybrids of Shigatoxigenic and Enterotoxigenic Escherichia coli (STEC/ETEC) Among Human and Animal Isolates in Finland

Diarrhoeagenic Escherichia coli (DEC) cause serious foodborne infections in humans. Total of 450 Shigatoxigenic E. coli (STEC) strains isolated from humans, animals and environment in Finland were examined by multiplex PCR targeting the virulence genes of various DEC pathogroups simultaneously. One per cent (3/291) of the human STEC and 14% (22/159) of the animal and environmental STEC had genes typically present in enterotoxigenic E. coli (ETEC). The strains possessed genes encoding both Shiga toxin 1 and/or 2 (stx₁ and/or stx₂) and ETEC‐specific heat‐stable (ST) enterotoxin Ia (estIa). The identified stx subtypes were stx_1a, stx_1c, stx_2a, stx_2d and stx_2g. The three human STEC/ETEC strains were isolated from the patients with haemolytic uraemic syndrome and diarrhoea and from an asymptomatic carrier. The animal STEC/ETEC strains were isolated from cattle and moose. The human and animal STEC/ETEC strains belonged to 11 serotypes, of which O2:H27, O15:H16, O101:H‐, O128:H8 and O141:H8 have previously been described to be associated with human disease. Identification of multiple virulence genes offers further information for assessing the virulence potential of STEC and other DEC. The emergence of novel hybrid pathogens should be taken into account in the patient care and epidemiological surveillance.     

Aerobic bacteriological studies on the respiratory tracts of apparently healthy and pneumonic camels (<Emphasis Type="Italic">Camelus dromedaries</Emphasis>) in selected districts of Afar Region,Ethiopia

A cross-sectional study was conducted to isolate and identify bacterial species from the respiratory tract of apparently healthy and pneumonic camels in Asayita and Dubti woredas in the Afar Region, Ethiopia. From a total of 74 lung tissue and 74 tracheal swab samples Staphylococcus aureus, 16.3%, Streptococcus equi subsp. equi, 13.0%, and Pasteurella multocida, 10.9%, were dominant isolates from pneumonic lungs; Escherichia coli, 12.7%, Proteus species, 10.9%, and Klebsiella pneumoniae, 9.1%, were the majority in the normal lungs. The majority of the isolates colonized both anatomical sites investigated. There was a statistically significant association between the health status of the camels as well as the anatomical site studied with the isolation rates of the major respiratory pathogens (p?<?0.05). Furthermore, the isolates were susceptible to norfloxacin, streptomycin, and gentamicin but resistant to ampicillin and tetracycline on in vitro test. Further studies on the pathogenicity of the major isolates are recommended.     

Prevalence,molecular fingerprinting and drug resistance profile of enterovirulent <Emphasis Type="Italic">Escherichia coli</Emphasis> isolates from free-ranging yaks of Tawang district,Arunachal Pradesh,India

Of 273 samples (rectal swab) collected from free-ranging yaks of Tawang district, Arunachal Pradesh, 42 Shiga toxin-producing Escherichia coli (STEC), six enteropathogenic E. coli (EPEC) and 27 enterotoxigenic E. coli (ETEC) strains were isolated. All the STEC and EPEC strains were further investigated for respective stx variants (for STEC only) and additional putative virulence factors. The 27 ETEC strains were also screened for characteristic enterotoxin gene(s) and colonization factors. Occurrence of ETEC was significantly (p < 0.05) higher in the diarrheic yaks and yaks of less than 1 year of age. Majority of enterovirulent E. coli isolates were resistant to amikacin, azithromycin, chloramphenicol, colistin, doxycycline, furazolidone, nalidixic acid, nitrofurantoin, streptomycin and tetracycline. Dendrogram, constructed with molecular fingerprinting profiles obtained from RAPD (Randomly Amplified Polymorphic DNA) and ERIC (Enterobacterial Repetitive Intergenic Consensus) PCR, placed the isolates in different clusters irrespective of their serotypes, virulence gene and drug resistance pattern. Collectively, the study indicates that yaks, being a potential reservoir of multidrug resistant STEC and EPEC, may represent significant risk to public health in this region. Higher recovery of ETEC isolates from yaks with diarrhea points out that ETEC may be a major determinant for repeated occurrence of diarrhea in yaks.     

Antibiotic resistance and molecular characteristics of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolated from backyard-raised pigs and pig workers

Staphylococcus aureus is a commensal and pathogenic bacterium with impact on public health and livestock industry. The study investigated nasal carriage, antibiotic resistance, and molecular characterization of S. aureus in pigs and pig workers. Nasal swabs from 300 backyard-raised pigs and 101 pig workers were used for the study. Resulting isolates were confirmed using MALDI-TOF MS, tested for antibiotic resistance, and three different multiplex PCRs were used to detect enterotoxin, mecA, spaA, scn, and pvl genes. spa typing was used to annotate the isolates into MLST clonal complexes (CC). Structured questionnaire was used to access possible risk factors for S. aureus carriage. The prevalence of S. aureus in pigs and pig workers were 5.3 and 12.9%, respectively. The isolates were resistant to beta-lactams (97%), tetracycline (62%), sulfonamide (52%), aminoglycoside (20.6%), fluoroquinolone (24%), and mupirocin (3.4%). Twenty seven (93%) of the isolates carried scn, 7(24%) pvl, and 12 (41%) enterotoxin genes, respectively. Questionnaire survey showed medical-related occupation of household members was associated (p?<?0.5) with S. aureus carriage. This study suggests the presence of human multidrug resistant strains of S. aureus, high carriage of pvl, and enterotoxin genes, and CC5, CC15, and CC152 were the CC-groups shared among pigs and pig workers.     

Antimicrobial resistance and virulence profile of enterococci isolated from poultry and cattle sources in Nigeria

This study investigated the occurrence, antimicrobial resistance and virulence of Enterococcus from poultry and cattle farms. Three hundred and ninety samples: cloacal/rectal swabs (n?=?260) and manure (n?=?130] were processed for recovery of Enterococcus species. Standard bacteriological methods were used to isolate, identify and characterize Enterococcus species for antimicrobial susceptibility and expression of virulence traits. Detection of antibiotic resistance and virulence genes was carried out by polymerase chain reaction. Enterococcus was recovered from 167 (42.8%) of the 390 samples tested with a predominance of Enterococcus faecium (27.7%). Other species detected were Enterococcus gallinarum, Enterococcus faecalis, Enterococcus hirae, Enterococcus raffinosus, Enterococcus avium, Enterococcus casseliflavus, Enterococcus mundtii and Enterococcus durans. All the isolates tested were susceptible to vancomycin, but resistance to tetracycline, erythromycin, ampicillin and gentamicin was also observed among 61.0, 61.0, 45.1 and 32.7% of the isolates, respectively. Sixty (53.1%) of the isolates were multidrug resistant presenting as 24 different resistance patterns with resistance to gentamicin-erythromycin-streptomycin-tetracycline (CN-ERY-STR-TET) being the most common (n?=?11) pattern. In addition to expression of virulence traits (haemolysin, gelatinase, biofilm production), antibiotic resistance (tetK, tetL, tetM, tetO and ermB) and virulence (asa1, gelE, cylA) genes were detected among the isolates. Also, in vitro transfer of resistance determinants was observed among 75% of the isolates tested. Our data revealed poultry, cattle and manure in this area are hosts to varying Enterococcus species harbouring virulence and resistance determinants that can be transferred to other organisms and also are important for causing nosocomial infection.     

Identification of bacteria present in ulcerative stomatitis lesions of captive sea turtles <Emphasis Type="Italic">Chelonia mydas</Emphasis>

Anthropogenic activities, predation, and diseases have contributed to a decrease in the sea turtle population in recent years. Ulcerative stomatitis is a condition that occurs in both wild and captive populations. The etiology of this condition is associated with bacteria such as E. coli, Citrobacter diversus, Klebsiella spp., Pseudomonas spp., Flavobacter calcoaceticus, Staphylococcus spp., and Flavobacterium spp. Some of these microorganisms are part of the oral microbiota of turtles, but alterations in the immune response can disturb the homeostatic relationship and cause an increase in the population of microorganisms, which in turn can cause disease. This work presents results on the isolation and identification of bacteria present in ulcerative stomatitis lesions in captive C. mydas turtles. Oral mucosa samples from 20 clinically healthy turtles and ten animals with ulcerative stomatitis lesions were studied. The samples were cultivated in enriched and differential media, and the identification was made using an automated method. The results showed a great diversity of bacteria in animals with ulcerative stomatitis with a higher prevalence of S. lentus and C. braakii was higher (60 and 50%, respectively) than in healthy animals. E. faecium was identified in 40% of diseased animals and 55% healthy animals. Turtles in this study had a diverse oral microbiota, and S. lentus and C. braakii may be involved in the etiopathogenesis of ulcerative stomatitis.     

Frequency of enterotoxins,toxic shock syndrome toxin-1, and biofilm formation genes in <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolates from cows with mastitis in the Northeast of Brazil

Staphylococcus aureus is among the microorganisms more frequently associated with subclinical bovine mastitis. S. aureus may produce several virulence factors. This study aimed at determining the frequency of virulence factors such as enterotoxins, toxic shock syndrome toxin 1, and ica adhesion genes. In addition, we assessed antimicrobial drug resistance in S. aureus isolated from clinical and subclinical cases of mastitis. A total of 88 cows with clinical or subclinical mastitis were sampled, resulting in 38 S. aureus isolates, from which 25 (65.78%) carried toxin genes, including seb, sec, sed, tst, and icaD adhesion gene. These S. aureus isolates belong to 21 ribotypes and three S. aureus strains belonged to the same ribotype producing ica adhesion gene. Approximately 90% of S. aureus strains obtained in our study demonstrated multiple resistance to different antimicrobial agents. The most efficacious antimicrobial agents against the isolates were gentamicin, amoxicillin, and norfloxacin. Gentamicin was the most efficacious agent inhibiting 78.95% of the S. aureus isolates. The least efficacious were penicillin, streptomycin, and ampicillin. Our results can help in understanding the relationship between virulence factors and subclinical mastitis caused by S. aureus. Further research about diversity of S. aureus isolates and genes responsible for the pathogenicity of subclinical mastitis is essential.     

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.     

Non‐O157 Shiga Toxin‐producing Escherichia coli Isolated from Diarrhoeic Calves in Argentina

Faecal samples from 76 diarrhoeic calves belonging to 36 farms located in the Pampas plain, Argentina, were examined for Shiga toxin‐producing Escherichia coli (STEC). A total of 15 STEC strains were isolated from 12 (15.8%) calves which came from six different farms. All stx positive strains assayed by PCR were also positives in the Vero cell cytotoxicity test. The majority (60.0%) of the STEC strains carried the stx₁ gene. Twelve (80.0%) of the STEC isolates which belonged to serotypes O5:H‐ (n = 4), O26:H11 (n = 4), O26:H‐ (n = 1), O111:H‐ (n = 2), and O123:H38 (n = 1) were also enterohaemolysin (EHly) positive and carried the gene encoding for intimin (eae). All the stx positive strains were negative for the bfpA gene. Localized adherence to HEp‐2 cells were observed in 83.3% of the eae+ STEC strains. STEC belonging to serotype O5:H‐ showed atypical biochemical properties, including urease production. Urease was also produced by two strains belonging to serotypes O153:H? and non‐typeable, respectively. Resistance to three or more antibiotics was observed in 12 (80.0%) of the STEC isolates. Most of the serotypes of STEC recovered in this survey carried virulence traits that are associated with increased human and bovine pathogenicity. The present study shows that highly virulent STEC strains are being shed by diarrhoeic calves from farms located in a high incidence area of human STEC infections.     

Shiga toxin-producing Escherichia coli O157:H7 from extensive cattle of the fighting bulls breed

Shiga toxin-producing Escherichia coli (STEC) O157:H7 represents a major public health concern worldwide, with cattle recognized as their main natural reservoir. The aim of this work was to determine the prevalence and the pheno-genotypic characteristics of STEC O157:H7 in a herd with 268 cattle of the fighting bulls breed (De Lidia breed) managed under extensive conditions in the South-West of Spain. Rectal-anal swabs of all animals were collected and examined for STEC O157:H7 by performing an immunomagnetic concentration and separation procedure combined with PCR, and the resulting isolates were characterized by both phenotypic and genotypic methods. Overall, STEC O157:H7 was isolated from seven animals (2.6%) in the herd. The PCR procedure indicated that all seven isolates displayed stx₂, eae-γ1, ehxA, O157 rfbE, and fliCh7 genes. They belonged to phage types 4 (one isolate) and 42 (two isolates), and four isolates reacted with typing phages but did not conform to a recognized pattern. Among the seven isolates there were five indistinguishable PFGE patterns and other two which differed only in ?2 restriction fragments, supporting the existence of horizontal transmission among animals in the herd. The present study demonstrates that cattle managed under extensive conditions in Spain can excrete STEC O157:H7 with their faeces. To our knowledge this is the first isolation of this pathogen from De Lidia cattle.     

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.     

Phylogenetic background and virulence genes of Escherichia coli isolates from colisepticemic and healthy broiler chickens in Iran

The purposes of this study were to determine the phylogenetic background and the virulence gene profiles of Escherichia coli isolates from colisepticemic and feces of healthy (AFEC) broiler chickens. In this study, 253 E. coli isolates including 141 avian pathogenic E. coli (APEC) and 112 AFEC isolates were examined by PCR. In general, 253 E. coli isolates distributed among group A (51.8%), B1 (15.8%), B2 (8.7%), and D (23.7%). Ten (8.9%) AFEC isolates segregated in to B1 phylo-group and 102 (91.1%) isolates fell into six different phylogenetic subgroups. Distribution of colisepticemic and fecal isolates differed significantly in their assignments to A and B1 phylo-groups. The three most prevalent virulence genes were crl, fimH, and aer in isolates between both groups. The four genetic markers aer, papC, afa, and sfa were detected significantly more often among colisepticemic isolates than in fecal isolates from healthy broilers. The presence of stx ₂ gene in fecal isolates were significantly differs among the colisepticemic isolates. F17 fimbrial family encoding gene and eae gene were detected in APEC and AFEC isolates, respectively. The colisepticemic and fecal isolates possessed the virulence genes were detected in all of the four phylogenetic groups. Several combination patterns of the virulence genes were detected in APEC and AFEC isolates. In colisepticemic isolates the combination of aer, crl, and fimH genes was the most prevalent pattern. None of the examined isolates harbored the cdt, cnf1, ipaH, and stx ₁ virulence gene sequences.