Occurrence of methicillin-resistant staphylococci in the pig-production chain in Ibadan,Nigeria

Staphylococcus species colonises humans and animals and is a major food contaminant with public health significance. Here, we assessed the occurrence of methicillin-resistant staphylococci (MRS) in the pig-production chain in Ibadan, Nigeria. Nares of 120 pigs and 10 farmers were sampled with sterile swabs whilst 54 pork samples were collected from a retail slaughterhouse. Staphylococcus species were isolated using enrichment, cefoxitin–aztreonam selective broth and Mannitol salt agar. Isolates were tested for susceptibility to cefoxitin (30 μg), oxacillin (1 μg) and vancomycin (30 μg). Methicillin-resistant staphylococci isolates were characterised using conventional biochemical tests. From 184 samples, 364 staphylococcal isolates were obtained. Amongst the 54 pork samples, 44.0% were contaminated with Staphylococcus species. Overall, 9 (2.5%) MRS were obtained and presumptively identified as Staphylococcus xylosus (n = 3), Staphylococcus sciuri (n = 3), Staphylococcus warneri (n = 2) and Staphylococcus cohnii (n = 1). There was no relationship between the prevalence of MRS between pigs and pig handlers in the farms, but Farm 2 had the highest frequency of 66.7% (p < 0.05). Piglets had the highest prevalence of 66.7% (p < 0.05) whilst MRS was absent in workers and pork samples. This study raises concerns about the cross-contamination of staphylococci in the food chain. Constant surveillance is imperative to ensure food safety.     

Occurrence of enterotoxigenic strains of Staphylococcus aureus in raw poultry meat

The aim of this work was to determine the contamination of raw poultry meat with enterotoxigenic strains of S. aureus, using the PCR method. PCR is a rapid and sensitive method, which can show the presence in food of enterotoxigenic strains of S. aureus on the basis of specific gene sequences and detect the potential source of contamination before enterotoxins are produced. No coagulase-positive staphylococci strains were found in 65 samples of chicken parts, but these bacteria were present in 11 of 23 examined samples of minced turkey meat (48%). Using the primers for enterotoxin genes A to C, 4 of the 11 isolated S. aureus strains showed a positive result in the PCR. Three of the isolates represented the SEB gene and remaining one the SEC gene. The results obtained showed that PCR is sensitive and rapid method which may be used for detection and identification of enterotoxigenic S. aureus.     

宁夏两家猪场猪萨佩罗病毒的检测与多样性分析

为了解宁夏地区猪群中萨佩罗病毒(PSV)的感染流行情况,本研究通过RT-PCR的方法对不同区域的2个规模化养猪场的160份临床健康猪咽拭子和肛拭子样品进行了检测。试验表明,所检的2个养殖场均存在PSV感染,总体平均阳性率为61.25%,其中咽拭子检出率为7.5%,肛拭子猪检出率为58.75%。试验结果表明PSV在宁夏地区普遍存在。     

Enterotoxin production by strains of Staphylococcus aureus isolated from animals and main in Nigeria

Enterotoxin production by 194 strains of Straphylococcus aureaus isolated from animals and man was investigated. The organisms were also classified into biotypes, irrespective of sources. Human biotypes isolated from people had the highest proportion of enterotoxigenic strains (39%). The animal biotypes isolated from human beings were not enterotoxinenic, but 8.8% of the human biotypes isolated from animals yielded enterotoxin. About 18% of the animal biotypes isolated from animals were enterotoxigenic, while 25% of the untypable biotypes from all the sources investigated were enterotoxin producers. The predominant enterotoxins were A and B, with A accounting for 55%5% of the entrotoxins produced by human biotypes. Enterotoxins A, B and C were produced by some animal biotypes, and three canine biotypes produced enterotoxin combinations, BE, CE and ABE, respectively. From goat isolates, only four of the caprine biotypes were enterotoxigenic, including the only isolate producing enterotoxins B, C and E. The isolates obtained from horses and chickens were non-enterotoxigenic regardless of the biotypes. It is concluded that strains producing enterotoxins A and B may be responsible for outbreaks of staphylococcal food poisoning in Nigeria. The close association between man and his animals in this environment may account for the high prevalence (47%) of human biotypes in animals.     

集约化养猪场嗜流产衣原体病流行状况的初步调查

本试验对北京郊区5个规模化猪场断奶仔猪、育成猪、育肥猪和怀孕母猪采集血清共507份,同时采集密切接触饲养人员、仔猪、育肥猪的喉头拭子、母猪阴道拭子及公猪的精液共183份。分别使用间接血凝诊断试剂盒、法国ELISA试剂盒检测抗体;利用直接荧光染色法检测抗原,包括166份猪喉头拭子、阴道拭子和精液以及17份饲养人员喉头拭子。结果发现间接血凝诊断试剂盒、ELISA试剂盒抗体阳性率分别为4.14%和2.17%;抗原平均阳性率为14.8%,其中精液阳性率达到37.5%,母猪阴道拭子阳性率为27.5%,饲养人员阳性率为23.5%。本试验证实北京郊区猪嗜流产衣原体在种公猪、母猪群和饲养员呈高流行趋势。同时研究结果显示,5个被调查的规模化猪场嗜流产衣原体抗体阳性率显著低于有报道的其他省市,这与间接血凝诊断试剂盒检测结果高于ELISA试剂有关。针对发现的问题,必须采取有效措施控制种公猪精液污染衣原体现象,阻断向母猪和饲养人员传播,以降低人兽共患病发生的风险。     

Assessment of the aerobic faecal microflora in mink (Mustela vison Schreiber) with emphasis on Escherichia coli and Staphylococcus intermedius

The present study was undertaken to investigate the culturable aerobic faecal microflora of mink from newborn until adulthood with emphasis on the potential pathogens Escherichia coli and beta-haemolytic coagulase positive staphylococci. Rectal swabs were taken from 10 healthy dams and their offspring on seven mink farms throughout the production season and a semi-quantitative enumeration of total E. coli and haemolytic E. coli, beta-haemolytic streptococci, beta-haemolytic coagulase positive staphylococci, total lactic acid bacteria, and enterococci was carried out in all samples using selective and non-selective media.Aerobic bacteria were cultured from close to 100% of the samples throughout the survey. Prevalence of E. coli isolates varied between 70 and 90% of the samples throughout the survey with a small decline at the end of the study period. The highest bacterial counts were found among recently weaned kits or kits in the early growth period (P<0.0012). Lactic acid bacteria and enterococci were isolated from more than 90% of all samples, while beta-haemolytic staphylococci were isolated from 20 to 70% of the samples. While beta-haemolytic staphylococci were dominant from birth and during the nursing period, counts of staphylococci gradually decreased during the nursing period and were outnumbered by E. coli during the growth season.     

Exudative Epidermitis in Pigs. Bacteriological Studies on the Causative Agent Staphylococcus Hyicus

The properties of six pathogenic and six non-pathogenic strains of staphylococci from clinical cases of exudative epidermitis are described. The pathogenic strains were shown to produce typical lesions following experimental inoculation of pigs. All of the strains studied were classified among the staphylococci due to their ability to produce acid from glucose under anaerobic conditions. One of the six pathogenic strains was a slow coagulase producer. It is suggested that the species designation Staphylococcus hyicus be retained.     

Enterotoxigenicity of staphylococci isolated from raw milk obtained from settled and nomadic herds around Zaria, Nigeria

Staphylococcal isolates from 135 raw milk samples obtained from settled herds (42) and nomadic herds (93), were characterized and assayed for enterotoxin production. Of the 42 samples from settled herds, 13 (31%) were California Mastitis Test (CMT)-positive, but all the samples contained staphylococci. Only 3 (3.2%) of the 93 raw milk samples from nomadic herds were CMT-positive but 58 (62.4%) contained staphylococci. Of the 13 isolates from CMT-positive milk obtained from settled herds, one produced enterotoxin A, while amongst the 29 from CMT-positive milk, 4 elaborated enterotoxin A and 3 produced type D. None of the isolates from milk obtained from nomadic herds was enterotoxigenic.     

Molecular characterization of Mycobacterium tuberculosis complex (MTBC) isolated from cattle in northeast and northwest China

We studied throat swabs and corresponding serum samples collected from 1067 protein purified derivative (PPD)-tuberculin skin test (TST) positive cattle from different regions of China. The 1067 throat swabs were inoculated onto modified Löwenstein–Jensen medium for the isolation and culture of Mycobacteria. Acid-fast bacilli were identified using traditional biochemical methods, polymerase chain reaction (PCR) amplification and multiplex PCR. They were distinguished as Mycobacterium tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) strains. An indirect Enzyme-Linked Immunosorbent Assay (ELISA) was applied to detect specific antibodies against bovine TB (bTB). Correlations among the ELISA, bacteriology and TST were analyzed and compared. Spoligotyping and variable number tandem repeats–mycobacterial interspersed repetitive unit (VNTR–MIRU) analysis were used to genotype the MTBC. In total, 111 strains of Mycobacteria were cultured from the 1067 throat swab samples, including 43 stains of MTBC (14 strains of Mycobacterium bovis and 29 of Mycobacterium tuberculosis) and 68 strains of NTM. Thirty-eight MTBC strains and four NTM strains were isolated from 72 throat swab samples that the ELISA determined were antibody positive; five MTBC strains and 64 NTM strains were isolated from 995 throat swab samples that were antibody negative on the ELISA. The positive isolation rates of MTBC and NTM were 38.7% (43/111) and 61.3% (68/111), respectively. The concordance rate of cultured MTBC with a positive result on the indirect ELISA for antibody was 52.8% (38/72), which was much higher than the positive rate for TST (4.0%; 43/1067). Genotyping of the 43 strains of MTBC isolated, using spoligotyping and VNTR–MIRU, showed that the 43 isolates had 26 genotypes; 16 strains had a unique genotype. Two groups of six strains and two strains, respectively, showed the same spoligotyping pattern, and belonged to the Beijing family and Beijing-like family, respectively. Combined application of spoligotyping and VNTR–MIRU typing would improve the molecular epidemiological investigation and monitoring of the etiology of bTB in China.     

Mycoplasma gallisepticum as a model to assess efficacy of inhalant therapy in budgerigars (Melopsittacus undulatus).

One hundred budgerigars (Melopsittacus undulatus) were infected in the nares (0.02 ml/naris), eye (0.02 ml/eye), and throat (0.2 ml) with Mycoplasma gallisepticum (MG) R strain (3.175 x 10(7) colony-forming units/ml). Fifty birds were treated with sterile broth and served as the controls; 25 of those were exposed to an inhalant, and the others were not treated. Infected birds were divided into four groups; 1) no treatment, 2) penicillin in drinking water, 3) inhalant, 4) both penicillin and inhalant. At days 3, 7, 10, 14, and 21 postinfection (PI), birds were assessed for clinical signs of disease. Five birds in each group were euthanatized at each interval PI; quantitative cultures were performed on lavages from the nares and trachea and on throat swabs; tracheas and air sacs were examined histopathologically. No clinical signs, lesions, or cultural isolations occurred in any control birds. All infected birds developed clinical signs and lesions of the trachea and air sac, but none died. The most severe clinical signs were seen in birds that were infected with MG and received no other intervention or birds that received penicillin in conjunction with infection. Increased respiratory tract lesions were associated with penicillin treatment; aerosol therapy resulted in fewer lesions.     

Biochemical properties and differentiation of coagulase-positive staphylococci from rooks and gulls

Coagulase-positive staphylococci were found in the throats of 46 rooks (69 per cent) and 47 gulls (21 per cent) out of totals of 67 and 229 birds, respectively. Of 111 strains isolated from throat swabs 86 (77.5 per cent) were classified as Staphylococcus aureus and 25 strains (22.5 per cent) as Staphylococcus intermedius. Of the S aureus strains 82 per cent from rooks and 50 per cent from gulls were biotyped. Most of them were identified as biotypes D and B, only a few as biotype A, and none as biotype C. Moreover, among the 24 S intermedius strains (96 per cent) biotypes 1 and 2 could be differentiated according to the type of growth on crystal violet agar and their ability to produce acetoin and acid from maltose, mannitol and trehalose.     

Occurrence, species distribution, antimicrobial resistance and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of domestic animals

beta-Lactams and macrolides are important antibiotics for treatment of staphylococcal infections in both humans and animals. The aim of the study was to investigate the occurrence, species distribution and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of dogs, horses, pigs, and cattle in Denmark. Nasal swabs were collected from a total of 400 animals, including 100 individuals of each species. Methicillin- and erythromycin-resistant staphylococci were isolated on selective media, identified by 16S rDNA sequencing, and typed by pulsed field gel electrophoresis (PFGE). Methicillin-resistant coagulase-negative staphylococci (MRCoNS) harbouring mecA were isolated from horses (50%) and dogs (13%), but not from food animals. The species identified were S. haemolyticus (n=21), S. vitulinus (n=19), S. sciuri (n=13), S. epidermidis (n=8), and S. warneri (n=2). mecA-mediated methicillin resistance in S. vitulinus was described for the first time. Methicillin-resistant S. aureus was not detected. PFGE analysis revealed the presence of specific MRCoNS clones in samples originating from the same veterinary hospital or equine farm. Erythromycin-resistant S. aureus (ERSA) was detected in 38% of pigs and all isolates harboured a constitutively expressed erm(C) gene. The vast majority (37/38) of pigs carrying ERSA originated from a farm characterized by frequent use of macrolides. Most ERSA isolates (28/38) displayed indistinguishable or closely related PFGE patterns, indicating clonal distribution within the farm. Based on the analysis of data on antimicrobial consumption, the occurrence of MRCoNS in companion animals and that of ERSA in pigs reflected national and local patterns of antimicrobial usage.     

Prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma

Currently, antimicrobial-resistant staphylococci, particularly methicillin-resistant Staphylococcus pseudintermedius (MRSP), are frequently isolated from canine superficial pyoderma in Japan. However, little is known regarding the nasal prevalence of MRSP in pet dogs. Here, we determined the prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma. Of the 125 nares and 108 affected sites of pet dogs with superficial pyoderma, 107 (13 species) and 110 (eight species) staphylococci strains, respectively, were isolated. The isolation rate of S. pseudintermedius from pyoderma sites (82/110 strains, 74.5%) was significantly higher than that from nares (57/107 strains, 53.3%) (P<0.01). Notably, the prevalence of MRSP (18/57 strains, 31.6%) in nares was equivalent to that in pyoderma sites (28/82 strains, 34.1%). Furthermore, the phenotypes and genotypes of antimicrobial resistance in MRSP strains from nares were similar to those from pyoderma sites. Our findings revealed that the prevalence of antimicrobial-resistant staphylococci in the nares of pet dogs with superficial pyoderma is the same level as that in affected sites. Therefore, considerable attention should be paid to the antimicrobial resistance of commensal staphylococci in companion animals.     

The prevalence of methicillin-resistant staphylococci in healthy horses in the Netherlands

Two hundred healthy horses housed at 23 different farms and one clinic and 42 persons in close contact with these horses were screened for the presence of methicillin resistant staphylococci. Samples were taken from the nose and the pastern of the horses and from the nose and throat of the humans and incubated in selective media. Isolates were identified by standard techniques and their susceptibilities were tested using an agar diffusion method. Methicillin-resistant strains were tested for the presence of the mecA gene by PCR. In 45 horses (22.5%) and 15 humans (35.7%) mecA positive staphylococci were found. All isolates were coagulase negative staphylococci, except for one methicillin-resistant Staphylococcus aureus isolated from a veterinarian. Staphylococcus sciuri was the predominant species found among the methicillin resistant staphylococci (MRS) in the horses, whereas S. epidermidis predominated in the humans. From the horses, often more than one species of MRS could be isolated, resulting in a total of 175 mecA positive equine isolates. The equine isolates were predominantly susceptible to most antimicrobials tested, whereas the human isolates showed more resistance. In conclusion, no methicillin-resistant Staphylococcus aureus was found in healthy horses in the Netherlands, but methicillin-resistant coagulase negative staphylococci were found frequently. Further studies are needed in order to investigate whether horses can be a reservoir for MRS or the mecA gene for humans.     

Distribution, characterization and genetic bases of erythromycin resistance in staphylococci and enterococci originating from livestock

The occurrence of staphylococci and enterococci expressing increased resistance to erythromycin (ERY) and, in particular, to macrolide-lincosamide-streptogramin B (MLS(B) ) antibiotics was investigated in dairy cattle, pigs and turkeys. Three hundred rectal (cloacal) swabs of each animal species were examined. A total of 120 and 71 staphylococci and enterococci, respectively, with increased resistance to ERY were identified. These were most frequent in turkeys (42.3% of positive animals), followed by pigs and dairy cattle (6.7% and 6.0% of positive animals, respectively). Similarly, MLS(B) -resistant isolates colonized predominantly turkeys (29.7% of animals), while their occurrence in pigs and dairy cattle was only sporadic (0.8% of animals). At least one of the erm genes encoding for MLS(B) resistance was found in 56.7% and 69.0% of staphylococci and enterococci, respectively. The erm(C) gene prevailed in staphylococci while the erm(B) gene was predominant in enterococci. Macrolide efflux genes msr(A) and msr(C) were also frequent in staphylococci and enterococci, respectively. Macrolide inactivation gene mph(C) occurred mainly in staphylococci. In staphylococci, methicillin resistance was rarely detected (7.5% of isolates), but resistance to telithromycin (ketolides) was frequent in both staphylococci and enterococci (89.2% and 47.9% of isolates, respectively). This study showed that turkeys represent an important source of ERY (MLS(B) )-resistant cocci. In addition, resistance to ketolides was also frequent.     

In vitro susceptibility testing of meticillin-resistant and meticillin-susceptible staphylococci to mupirocin and novobiocin

Antimicrobials effective against meticillin-resistant staphylococci are limited. Mupirocin is a topical antimicrobial used to treat bacterial skin infections. Novobiocin is an oral antimicrobial approved for treatment of staphylococcal upper respiratory infections in dogs. This study reports the in vitro activity of mupirocin and novobiocin on meticillin-susceptible (MSS) and resistant staphylococci (MRS) from healthy dogs and dogs with superficial pyoderma. Staphylococci were isolated from skin swabs at four sites on healthy dogs and from lesions on dogs with superficial pyoderma. Staphylococci were identified by morphology and by catalase and coagulase testing. Speciation and susceptibility testing were performed by the Dade Microscan (W. Sacramento, CA, USA). Meticillin resistance was confirmed by an oxacillin screen plate. Novobiocin and mupirocin susceptibilities were tested by disc diffusion. Staphylococci were cultured from 61 healthy dogs (17 MRS and 44 MSS) and 30 dogs with pyoderma (15 MRS and 15 MSS), with higher proportions of MRS isolates in dogs with pyoderma (P=0.038; χ(2) test). For mupirocin, 79.5% (35 of 44) MSS and 82.3% (14 of 17) MRS isolates from healthy dogs, and 100% (15 of 15) MSS and 86.6% (13 of 15) MRS isolates from dogs with pyoderma were susceptible (MSS, P=0.094; MRS, P=1.0; Fisher's exact test). For novobiocin, 95.4% (42 of 44) MSS and 52.9% (nine of 17) MRS isolates from healthy dogs and 93.3% (14 of 15) MSS and 80% (12 of 15) MRS isolates from dogs with pyoderma were susceptible (MSS, P=1.0; MRS, P=0.148; Fisher's exact test).     

A bacteriologic study of scabby-hip lesions from broiler chickens in Texas

Broilers from commercial flocks experiencing a 10-60% incidence of scabby-hip lesions at processing were examined, and selected skin lesions were cultured. Over 70% of the lesions were associated with traumatic excoriations, particularly on the caudal dorsal convexity of the birds. Most lesions were observed on birds that were 5 weeks of age or older. From the 27 specimens cultured, Clostridium perfringens was isolated in pure culture from 4 lesions and Staphylococcus species from 10 lesions. Pure cultures of staphylococci were recovered from 4 lesions, and 2-5 different staphylococci were isolated from 6 lesions. Eight staphylococci were identified as S. sciuri, 8 as S. simulans, 2 as S. epidermidis, 2 as S. lentus, 2 as S. warneri, 1 as S. cohnii, and 1 as S. intermedius. Fifty cutaneous specimens from 10 5-week-old normal broilers were cultured. A total of 197 isolates were identified including 65 S. sciuri, 52 S. lentus, 24 S. simulans, 12 S. hyicus, 11 S. warneri, 9 S. cohnii, 9 S. gallinarium, 8 S. xylosus, and 7 S. epidermidis.     

Sheep pox disease outbreaks in Madras Red and Mechery breeds of indigenous sheep in Tamilnadu,India

Sheep pox disease outbreaks were recorded among Madras Red (n = 145) and Mechery (n = 80) breeds of indigenous sheep on three farms in Tamilnadu. Over both breeds, adult mortality rate ranged from 2.66% to 37.5% and lamb mortality ranged from 10% to 17.33%. However, mortality was more in Mechery sheep (50% overall; 37.5% adults, 12.5% lambs) than in Madras Red sheep (24.28% overall; 10.34% adults, 13.79% lambs). The clinical signs observed were high fever, anorexia, respiratory distress, mucopurulent nasal discharge and in a few cases diarrhoea. Cutaneous lesions were mainly observed around nostrils, eyes, lips, ears and in the abdomen. Most of the lesions were covered with purulent materials and on cleaning with sterile swabs, fresh wounds were observed. Dry scabs were also observed over the oral commissure and maxillary areas, which on removal exposed fresh wounds. Important observations on necropsy were severe nodular lesions in the lungs and intestine. The disease was diagnosed as sheep pox by agar gel immunodiffusion test, isolation of virus and its neutralization in BHK₂₁ cells by specific antiserum and by electron microscopy.     

Aerobic bacteria from mucous membranes, ear canals, and skin wounds of feral cats in Grenada, and the antimicrobial drug susceptibility of major isolates

In a 2-year period 54 feral cats were captured in Grenada, West Indies, and a total of 383 samples consisting of swabs from rectum, vagina, ears, eyes, mouth, nose and wounds/abscesses, were cultured for aerobic bacteria and campylobacters. A total of 251 bacterial isolates were obtained, of which 205 were identified to species level and 46 to genus level. A commercial bacterial identification system (API/Biomerieux), was used for this purpose. The most common species was Escherichia coli (N = 60), followed by Staphylococcus felis/simulans (40), S. hominis (16), S. haemolyticus (12), Streptococcus canis (9), Proteus mirabilis (8), Pasteurella multocida (7), Streptococcus mitis (7), Staphylococcus xylosus (7), S. capitis (6), S. chromogenes (4), S. sciuri (3), S. auricularis (2), S. lentus (2), S. hyicus (2), Streptococcus suis (2) and Pseudomonas argentinensis (2). Sixteen other isolates were identified to species level. A molecular method using 16S rRNA sequencing was used to confirm/identify 22 isolates. Salmonella or campylobacters were not isolated from rectal swabs. E. coli and S. felis/simulans together constituted 50% of isolates from vagina. S. felis/simulans was the most common species from culture positive ear and eye samples. P. multocida was isolated from 15% of mouth samples. Coagulase-negative staphylococci were the most common isolates from nose and wound swabs. Staphylococcus aureus, or S. intemedius/S. pseudintermedius were not isolated from any sample. Antimicrobial drug resistance was minimal, most isolates being susceptible to all drugs tested against, including tetracycline.     

A possible role of capybaras (Hydrochoerus hydrochoeris hydrochoeris) in foot-and-mouth disease (FMD) endemicity

The susceptibility of capybaras exposed to foot-and-mouth disease (FMD) virus by the intramuscular route and the rodents' close coexistence with cattle in FMD endemic ecosystems suggested that the species might play an important role in the virus' survival in the field.In the present study 2 capybaras and 2 cattle were exposed by contact to a capybara inoculated intramuscularly with FMD virus. Both pairs of exposed animals were then used as a contact source with another 2 cattle and 2 capybaras, respectively. All the animals became infected prior to the appearance of clinical lesions in the respective donor animals and developed generalized FMD clinical lesions. Specific neutralizing antibodies and antibodies to virus-infection-associated antigen (VIA) were also developed.Virus was isolated from feces and from throat swabs of 1 of the capybaras up to 17, but not at 23 days post-contact. Virus was isolated from the remaining animals up to 7–14 days post-contact.The results indicate that these rodents might transmit virus over long distances due to their migratory movements, but probably do not act as natural virus reservoirs.