Detection of Pathogenic Serogroups and Virulence Genes in Listeria monocytogenes Strains Isolated from Beef and Beef Products Retailed in Gauteng Province, South Africa, Using Phenotypic and Polymerase Chain Reaction (PCR)-Based Methods.

South Africa recently (2017-18) experienced the largest outbreak of human listeriosis in the world caused by L. monocytogenes following the consumption of "polony," a ready-to-eat meat product. Most (59%) cases originated from Gauteng province, South Africa. As a follow-up study to the outbreak, we used standard bacteriological and molecular methods to determine the prevalence of pathogenic and virulent serogroups of L. monocytogenes in various beef and beef products retailed in Gauteng province, South Africa. The overall prevalence of Listeria spp. was 28% (112/400), comprising Listeria monocytogenes (9.3%), Listeria innocua (16.3%), and Listeria welshimeri (2.5%) (p < 0.001). It is crucial to have detected that the region (p=0.036), type of product (p=0.032), and temperature at storage (p=0.011) significantly affected the occurrence of L. monocytogenes in beef products. It is alarming that pathogenic serogroups 4b-4d-4e (51.4%) and 1/2a-3a (43.2%) were detected among the isolates of L. monocytogenes. Importantly, they were all carriers of seven virulence-associated genes (hlyA, inlB, plcA, iap, inlA, inlC, and inlJ). Our study also demonstrated that 16.7% of "polony" samples investigated were contaminated with L. monocytogenes. Considering that pathogenic and virulent L. monocytogenes contaminated beef and beef products retailed in South Africa, the food safety risk posed to consumers remains and cannot be ignored. Therefore, it is imperative to reduce the contamination of these products with L. monocytogenes during beef production, processing, and retailing to avoid future outbreaks of human listeriosis in the country.

Absence of Tuberculosis-Causing Mycobacteria from Slaughtered Livestock Tissues and Environmental Samples, Gauteng Province, South Africa.

Mycobacterium tuberculosis complex (MTBC) is a group of bacteria responsible for causing tuberculosis in animals and humans. In South Africa (S.A), slaughterhouses are registered by the government and closely inspected and audited for hygienic slaughter practices. Meat inspection to detect lesions has been used for passive surveillance, monitoring, and diagnosis of the disease status. Information on the current status of bovine tuberculosis (bTB) in livestock in the country is limited. Hence, we investigated the occurrence of Mycobacterium spp. in the tissues of slaughtered livestock and environmental samples in abattoirs in Gauteng province of South Africa (S.A). The cross-sectional study employing random sampling from cattle, pigs, and sheep (with the collection of liver, lung, spleen, and different lymph nodes) irrespective of lesions was carried out in 19 red meat abattoirs. Five hundred animals were sampled, comprising cattle (n = 369), pigs (n = 90), and sheep (n = 41). Additionally, 19 environmental samples were collected from feedlots, or where animals drink water while awaiting slaughter, to identify mycobacterial species using culture, acid-fast bacteria staining, and polymerase chain reaction (PCR). The Chi-square and Fisher's Exact tests were used to detect statistically significant differences in the frequency of detection of Mycobacterium spp. according to the variables investigated (types of tissues, livestock, abattoirs, etc.). The PCR assays detected no MTBC complex species DNA in the bacterial isolates from cattle (n = 32). Sequence analysis (16S rDNA) of the isolates from eight cattle confirmed only two species, namely Mycobacterium colombiense (99.81% identity) and Mycobacterium simiae (99.42% identity). The remaining isolates were identified as members of the Actinomadura species. From the environmental samples, bacterial isolation was made from three samples, and two could only be identified up to the genus level (Mycobacterium species) while the remaining isolate was identified as Mycobacterium senuense (99.22% identity). The study revealed the absence of bovine tuberculosis-causing pathogens in red meat abattoirs of the Gauteng province. Although non-tuberculous Mycobacteria have been implicated as potentially causing tuberculosis-like diseases in livestock, their occurrence in the current study was found to be low, but the potential to cause disease cannot be ignored.

Brucellosis Seropositivity Using Three Serological Tests and Associated Risk Factors in Abattoir Workers in Gauteng Province, South Africa.

Abattoir workers are liable to zoonotic infections from animals and animal products, primarily to diseases with asymptomatic and chronic clinical manifestations in animals, such as brucellosis. No published reports exist on the seroprevalence of brucellosis in abattoir workers in South Africa. Therefore, this cross-sectional study was conducted to estimate the occurrence and risk factors for Brucella exposure in abattoir workers in Gauteng Province. A total of 103 abattoir workers and managers from 6 abattoirs, where brucellosis-positive slaughtered cattle and sheep were previously detected, were interviewed and tested with serological assays using the Rose Bengal test (RBT), BrucellaCapt, and IgG-ELISA. A pre-tested questionnaire was administered to consenting respondents to obtain information on risk factors for brucellosis. Of the 103 respondents tested, the distribution of female and male workers was 16 (15.5%) and 87 (84.5%), respectively. The seroprevalence for exposure to brucellosis was 21/103 (20.4%, 95%CI: 13.1-29.5) using a combination of RBT, BrucellaCapt, or IgG-ELISA. For test-specific results, seroprevalences by RBT, BrucellaCapt, and IgG-ELISA were 13/103 (12.6%, 95%CI: 6.9-20.6), 9/103 (8.74%, 95%CI: 4.1-15.9), and 18/103 (17.5%, 95%CI: 10.7-26.2), respectively. Low-throughput abattoirs were identified as associated risks, as 29.3% of workers were seropositive compared with 12.7% of workers in high-throughput abattoirs, which highlights that direct contact at abattoirs poses higher risk to workers than indirect and direct contact outside abattoirs. This study confirms the occurrence of Brucella spp. antibodies among abattoir workers in South Africa, possibly due to occupational exposure to Brucella spp., and highlights the occupational hazard to workers. Furthermore, findings underscore that abattoir facilities can serve as points for active and passive surveillance for indicators of diseases of public health importance. We recommend periodic implementation of brucellosis testing of abattoir workers country-wide to establish baseline data for informing appropriate preventive practices and reducing the potential burden of infection rates among these high-risk workers.

Q fever and toxoplasmosis in South African livestock and wildlife: a retrospective study on seropositivity, sporadic abortion, and stillbirth cases in livestock caused by Coxiella burnetii.

BACKGROUND: Q fever and toxoplasmosis are economically important zoonoses as they cause considerable losses in livestock (cattle, sheep and goats) and wildlife (antelopes, giraffes, lions, and cheetahs) through reproductive disorders such as abortions and stillbirths. Q fever and toxoplasmosis testing in South Africa is conducted by the Agricultural Research Council-Onderstepoort Veterinary Research (ARC-OVR). However, both zoonoses are understudied and not monitored in South Africa as they are not considered controlled or notifiable diseases in the Animal Disease Act 35 of 1984. A retrospective study was conducted on Q fever (2007-2009) and toxoplasmosis (2007-2017) using diagnostic laboratory data at the ARC-OVR. Also, we report on sporadic abortion and stillbirth cases in livestock from diagnostic tissue samples submitted for Coxiella burnetii polymerase chain reaction (PCR) detection at the ARC-OVR. RESULTS: During 2007 to 2009, 766 animal samples were tested for C. burnetii antibodies and seropositivity was 0.9% (95%CI: 0.3-1.7) with sheep (1.9%; 95%CI: 0.6-4.4) having the highest seropositivity followed by cattle (0.7%; 95%CI: 0.09-2.6), while all goats (0.0%; 95%CI: 0.0-4.2) and wildlife (0.0%; 95%CI: 0.0-2.5) tested were negative. From 2007 to 2017, 567 sera were tested for T. gondii antibodies; overall seropositivity was 12.2% (95%CI: 9.6-15). Wildlife had highest seropositivity to T. gondii antibodies (13.9%; 95%CI: 9.0-19.7) followed by goats (12.9%; 95%CI: 9.2-17.4) and sheep (12.3%; 95%CI: 5.1-23.8) while seropositivity in cattle was 2.4% (95%CI: 0.06-12.9). Of 11 animals tested by C. burnetii PCR detection (2021-2022), 10 (91.0%) were positive. The amplicon sequences showed similarity to Coxiella burnetii strain 54T1 transposase gene partial coding sequence. CONCLUSIONS: We have confirmed the occurrence of the causative agents of Q fever and toxoplasmosis in livestock and wildlife in South Africa, with data limitations. These zoonoses remain of importance with limited information about them in South Africa. This study provides baseline information for future studies on Q fever and toxoplasmosis in South African livestock and wildlife, as well other African countries. Due to limited data collection experienced in this study, it is recommended that improvements in data collection samples tested should include associated factors such as sex, age, and breed of the animals.

Prevalence and risk factors of Q fever (Coxiella burnetii) in cattle on farms of Limpopo province, South Africa.

Q fever in animals and humans and its economic and public health significance has been widely reported worldwide but in South Africa. There are few studies on the prevalence of this zoonosis and its associated risk factors in South African livestock. Therefore, a cross-sectional study was conducted to determine the seroprevalence, molecular prevalence, and risk factors associated with C. burnetii in cattle on farms in South Africa's Limpopo province. Out of 383 cattle tested for antibodies, the overall seroprevalence was 24.28%. Herd size of >150 (OR: 9.88; 95%CI: 3.92-24.89; p < 0.01) remained associated with C. burnetii seropositivity in cattle. For PCR detection, targeting IS1111 fragment, cattle with no abortion history (OR: 0.37; 95%CI: 0.18-0.77; p < 0.01) and herd size of >150 (OR: 3.52; 95%CI: 1.34-9.24; p < 0.01) remained associated with C. burnetii positivity. The molecular prevalence in sheath scrapings and vaginal swabs by IS1111 PCR was 15.67%. Cohen's kappa agreement test revealed a fair agreement between the PCR and ELISA results (k = 0.40). Sequence analysis revealed that the amplicons had similarities to the C. burnetii transposase gene fragment, confirming the presence of the pathogen. The higher seroprevalence than molecular prevalence indicated a past C. burnetii infection, no bacterial shedding through vaginal mucus in cows, or preputial discharge in bulls. Similarly, the detection of C. burnetii by PCR in the absence of antibodies could be partly explained by recent infections in which antibodies have not yet been produced against the bacteria, or the level of these antibodies was below the detectability threshold. The presence of the pathogen in cattle and the evidence of exposure, as shown by both PCR and ELISA suggests an active circulation of the pathogen. This study demonstrated that C. burnetii is widespread in the study area and that a herd size of >150 is associated with C. burnetii seroprevalence and molecular prevalence.

Molecular Characterization of Leptospira Species Detected in the Kidneys of Slaughtered Livestock in Abattoirs in Gauteng Province, South Africa.

Leptospira was investigated in kidneys (n = 305) from slaughtered livestock in the Gauteng Province abattoirs, South Africa, using a culture medium to isolate Leptospira, followed by the LipL32 qPCR to detect Leptospira DNA. The SecY gene region was amplified, sequenced, and analyzed for LipL32 qPCR-positive samples or Leptospira isolates. The overall frequency of isolation of Leptospira spp. was 3.9% (12/305), comprising 4.8% (9/186), 4.1% (3/74), and 0% (0/45) from cattle, pigs, and sheep, respectively (p > 0.05). However, with LipL32 qPCR, the overall frequency of Leptospira DNA was 27.5%, consisting of 26.9%, 20.3%, and 42.2% for cattle, pigs, and sheep, respectively (p = 0.03). Based on 22 SecY sequences, the phylogenetic tree identified the L. interrogans cluster with serovar Icterohaemorrhagiae and the L. borgpetersenii cluster with serovar Hardjo bovis strain Lely 607. This study is the first molecular characterization of Leptospira spp. from livestock in South Africa. The reference laboratory uses an eight-serovar microscopic agglutination test panel for leptospirosis diagnosis, of which L. borgpetersenii serovar Hardjo bovis is not part. Our data show that pathogenic L. interrogans and L. borgpetersenii are circulating in the livestock population. Diagnostic use of molecular methods will eliminate or reduce the under-reporting of leptospirosis in livestock, particularly sheep, in South Africa.

Phylogenetic analyses of Salmonella detected along the broiler production chain in Trinidad and Tobago.

This study was conducted to determine the phylogenies of Salmonella strains isolated from cross-sectional studies conducted at hatcheries, broiler farms, processing plants, and retail outlets (broiler production chain) in Trinidad and Tobago over 4 yr (2016-2019). Whole-genome sequencing (WGS) was used to characterize Salmonella isolates. Core genome phylogenies of 8 serovars of public health significance were analyzed for similarities in origin and relatedness. In addition, Salmonella strains isolated from human salmonellosis cases in Trinidad were analyzed for their relatedness to the isolates detected along the broiler production chain. The common source of these isolates of diverse serovars within farms, within processing plants, between processing plants and retail outlets, and among farm-processing plant-retail outlet continuum was well-supported (bootstrap value >70%) by the core genome phylogenies for the respective serovars. Also, genome analyses revealed clustering of Salmonella serovars of regional (intra-Caribbean) and international (extra-Caribbean) origin. Similarly, strains of S. Enteritidis and S. Infantis isolated from human clinical salmonellosis in 2019 from Trinidad and Tobago clustered with our processing plant isolates recovered in 2018. This study is the first phylogenetic analysis of Salmonella isolates using WGS from the broiler industry in the Caribbean region. The use of WGS confirmed the genetic relatedness and transmission of Salmonella serovars contaminating chickens in broiler processing, and retailing in the country, with zoonotic and food safety implications for humans.

Characterisation and epidemiological subtyping of Shiga toxin-producing Escherichia coli isolated from the beef production chain in Gauteng, South Africa.

In South Africa, there is a shortage of epidemiologic data on Shiga toxin-producing Escherichia coli (STEC) in the beef production chain. This study was conducted to characterise STEC isolates originating from three studies conducted in a cattle feedlot, beef abattoirs and retail outlets in Gauteng province, South Africa. Polymerase chain reaction was used to detect virulence genes, the Epsilometer test to assess antimicrobial susceptibility, pulsed-field gel electrophoresis (PFGE) to investigate genetic relatedness of isolates, and conventional serotyping for phenotypic identification. Amongst the 86 STEC isolates, the eaeA gene was detected in 20 (23%), and 26 different serogroups were identified, including the clinically important O8, O174, O2, 020 and O117. The majority of the isolates (95%; 82/86) exhibited resistance to one or more antimicrobial agents, and 30 of the isolates (35%) exhibited multi-drug resistance (MDR), being resistant to at least three antimicrobial classes. The PFGE patterns showed a highly diverse but related STEC population, with 45 distinct patterns and evidence of horizontal transmission along the beef production chain. This is significant because it demonstrates continual environmental contamination and risk of contamination along the beef production chain and the food chain. To our knowledge, this is the first study that provides evidence of horizontal transmission of STEC along the beef production chain in South Africa. This epidemiological information could facilitate the development of a proactive strategy for reducing potential foodborne outbreaks and transmission of antimicrobial resistant pathogens in the food chain.

Prevalence and Characterization of Campylobacter Species from Chickens Sold at Informal Chicken Markets in Gauteng, South Africa.

ABSTRACT: This study determined the prevalence, characteristics, and risk factors of Campylobacter species contamination of chicken carcasses sold at informal poultry outlets in Gauteng province, South Africa. Within six townships, 151 chicken carcasses were collected from 47 outlets. Carcass swab, cloacal swab, and carcass drip samples were collected from each chicken, along with a matched questionnaire on risk factors regarding Campylobacter contamination. Sample-inoculated Bolton broth (BB) was cultured to isolate Campylobacter species by bacteriological methods. Subsequent confirmation and characterization of Campylobacter were conducted using polymerase chain reaction (PCR). Isolated Campylobacter strains were evaluated for the presence of six virulence genes (ciaB, dnaj, pldA, racR, flaA, and flaB), three toxin genes (cdtA, cdtB, and cdtC), and one antimicrobial resistance gene (tetO). The overall prevalence of Campylobacter was 23.4% (106 of 453), with sample type-specific prevalence being 17.2% (26 of 151), 25.8% (39 of 151), and 27.2% (41 of 151) for the carcass swabs, cloacal swabs, and carcass drip, respectively, following bacteriological isolation and confirmation by PCR. The overall prevalence of Campylobacter species was 93.5% by PCR, which varied significantly (P = 0.000) by sample: 99.2, 98.4, and 82.8% for carcass swabs, cloacal swabs, and carcass drip, respectively, by using PCR to detect Campylobacter in BB. Important risk factors for carcass contamination by Campylobacter included the slaughter of culled breeders and spent chickens, the use of stagnant water, and poor sanitation. Virulence and toxin gene frequencies were higher in C. jejuni-positive (82.5%) than in C. coli-positive (71.4%) BB cultures, but tetracycline resistance gene (tetO) frequency was higher in C. coli (75.9%) than in C. jejuni (48.10%). The observed high frequencies in C. jejuni recovered from street-vended chickens may pose food safety and therapeutic concerns to consumers.

Genomic Comparison of Eight Closed Genomes of Multidrug-Resistant Salmonella enterica Strains Isolated From Broiler Farms and Processing Plants in Trinidad and Tobago.

Salmonella enterica is an important foodborne pathogen worldwide. We used long and short-read sequencing to close genomes of eight multidrug-resistant (MDR) S. enterica strains, belonging to serovars Infantis (2), Albany, Oranienburg, I 4,[5],12:i:-, Javiana, Schwarzengrund, and Kentucky from broiler chicken farms and processing plants in Trinidad and Tobago. They also belonged to seven different sequence types (STs- 32, 292, 1510, 19, 24, 152, and 96). Among the strains, seven had demonstrated multi-drug resistance with the presence of at least three AMR genes, whereas three isolates contained the quinolone resistance gene qnr B19 in plasmids (CFSAN103840, CFSAN103854, and CFSAN103872). The extended-spectrum ß-lactamase genes bla CTX-M-65 (CFSAN103796) and bla TEM-1 (CFSAN103852) were detected in this study. The genomes closed in this study will be useful for future source tracking and outbreak investigations in Trinidad and Tobago and worldwide.

Multiple-Locus Variable-Number Tandem Repeat Analysis Genotypes of Listeria monocytogenes Isolated from Farms, Abattoirs, and Retail in Gauteng Province, South Africa.

ABSTRACT: The use of multiple-locus variable-number analysis (MLVA) of tandem repeats (TRs) for subtyping Listeria monocytogenes has proven to be reliable and fast. This study determined the MLVA genotypes of 60 isolates of L. monocytogenes recovered from cattle farms, abattoirs, and retail outlets in Gauteng province, South Africa. The distribution of the 60 L. monocytogenes isolates analyzed by type of sample was as follows: raw beef (28, 46.7%), ready-to-eat beef products (9, 15.0%), beef carcass swabs (9, 15.0%), cattle environment (6, 10.0%), and cattle feces (8, 13.3%). The serogroups of the isolates were determined using PCR and the MLVA genotypes based on six selected loci. The frequency of the 60 serogroups detected was as follows: 1/2a-3a (IIa) (27, 45.0%); 4b-4d-4e (1Vb) (24, 40.0%); 1/2c-3c (IIc) (8, 13.3%); and 1/2b-3b (IIb) (1, 1.7%). MLVA successfully clustered genetically related isolates and differentiated nonrelated isolates, irrespective of their sources, sample types, and serogroups, as demonstrated by 16 MLVA pattern types detected. For serogroup 4b-4d-4e (IVb), there was no variation in TRs LM-TR2, LM-TR4, and LM-TR6, which each contained only one allele (02, 00, and 93, respectively). However, across the sources and sample types of isolates, there was variation in serogroup 4b-4d-4e (IVb): LM-TR1 contained 00, 03, and 05; LM-TR3 contained 14, 20, and 22; and LM-TR5 contained 14, 21, and 25. Similar patterns of variation in the TRs were detected in the other serogroups (1/2a-3a, 1/2b-3b, and 1/2c-3c). BioNumeric data analysis identified at least five types in Gauteng province. MLVA epidemiologically clustered the related isolates and differentiated unrelated isolates.

Occurrence, Risk Factors, Serotypes, and Antimicrobial Resistance of Salmonella Strains Isolated from Imported Fertile Hatching Eggs, Hatcheries, and Broiler Farms in Trinidad and Tobago.

ABSTRACT: This cross-sectional study was conducted to determine the occurrence, risk factors, and characteristics of Salmonella isolates recovered from imported fertile broiler hatching eggs, hatcheries, and broiler farms in Trinidad and Tobago. Standard methods were used to isolate and characterize Salmonella isolates from two broiler hatcheries and 27 broiler farms in the country. The frequency of isolation of Salmonella was 0.0% for imported fertile hatching eggs (0 of 45 pools of 10 eggs each, i.e., 450 eggs), 7.6% for hatcheries (12 of 158 samples), and 2.8% for broiler farms (24 of 866 samples) (P = 0.006). Stillborn chicks at hatcheries had the highest prevalence of Salmonella (7 of 28 samples, 28.0%), whereas on broiler farms the cloacal swabs had the highest prevalence of Salmonella (15 of 675 samples, 2.2%). None of the 15 farm management and production practices investigated were significantly associated (P > 0.05) with the isolation of Salmonella. The predominant Salmonella serotypes were Kentucky (83.3%) and Infantis (62.5%) among hatchery and farm isolates, respectively. The disk diffusion method revealed frequencies of antimicrobial resistance (i.e., resistance to one or more agents) of 44.0% (11 of 25 isolates) and 87.5% (35 of 40 isolates) at hatcheries and broiler farms, respectively (P = 0.0002). Antimicrobial resistance among hatchery isolates was highest (28.0%) to doxycycline and kanamycin and was very high (>65%) among farm isolates to sulfamethoxazole-trimethoprim, gentamicin, ceftriaxone, kanamycin, and doxycycline. Multidrug resistance (MDR; i.e., resistance to antimicrobial agents from three or more classes) was exhibited by 4.0 and 85.7% of Salmonella isolates recovered from several environmental and animal sources at the hatcheries and farms, respectively (P < 0.0001). The high level of antimicrobial resistance and the presence of MDR among Salmonella isolates from broiler farms highlight the therapeutic implications and the potential for MDR strains to enter the food chain.

Application of the gamma-interferon assay to determine the prevalence of bovine tuberculosis in slaughter livestock at abattoirs in Gauteng, South Africa.

BACKGROUND: Bovine tuberculosis (bTB) is a zoonotic disease with great economic impact estimated at billions of dollars annually worldwide. Meat inspection represents a long-standing form of disease surveillance that serves both food safety and animal health. The objective of this study was to determine the prevalence of bTB in livestock at abattoirs using a cell-mediated immune (CMI) assay, the gamma interferon (IFN-γ) assay. This cross-sectional study was conducted at selected abattoirs (low-throughput, high-throughput and rural/informal) in Gauteng province, where animals were also subjected to routine meat inspection. RESULTS: A total of 410 fresh blood samples were collected from slaughter livestock (369 cattle and 41 sheep) from 15 abattoirs, and analysed using Bovigam® test kit with bovine, avian and Fortuitum purified protein derivatives (PPD) as blood stimulating antigens. The estimated prevalence of bTB in cattle was 4.4% (95% CI: 2.4%-7.3%). The prevalence of bTB in cattle varied between abattoirs (p = .005), ranging from 0% to 23%; however, there were no significant differences among genders, breeds, municipality, districts, origins of animals (feedlot, auction or farm) or throughput of abattoirs. The prevalence of avian reactors was 6.0% (95% CI: 3.6%-9.2%) in cattle, varying between abattoirs (p = .004) and ranging from 0% to 20.7%. None of the sheep with valid test results was positive for bTB and none was avian reactors (95% CI: 0%-15%). CONCLUSION: The detection of bTB reactor cattle in our study clearly shows the limitation of disease surveillance using a meat inspection approach, as all the 410 slaughter animals sampled had passed visual abattoir inspection and been classified as bTB-free. Our findings therefore emphasize the risk of zoonotic transmission of bTB to abattoir workers and potential food safety hazard to consumers. Furthermore, our study highlights the potential for the use of the IFN-γ assay to reduce this risk.

Closed Genome Sequence of a Salmonella enterica Serotype Senftenberg Strain Carrying the mcr-9 Gene Isolated from Broken Chicken Eggshells in Trinidad and Tobago.

Salmonella enterica is a highly important foodborne pathogen worldwide. We report the complete genome sequence of a sequence type 14 Salmonella enterica serotype Senftenberg strain carrying the mcr-9 gene in a plasmid isolated from broken chicken eggshells in Trinidad and Tobago, obtained by using a combination of long- and short-read sequencing.

Antimicrobial Resistance Profiles of Salmonella Isolates on Chickens Processed and Retailed at Outlets of the Informal Market in Gauteng Province, South Africa.

The study determined the antimicrobial resistance profiles of Salmonella on chickens processed and retailed at outlets of the informal markets in Gauteng province, South Africa. The study also investigated the relationship of antimicrobial resistant Salmonella to the source and type of samples and their serotypes. Carcass swabs, cloacal swabs and carcass drips were randomly collected from each of 151 slaughtered chickens from six townships. Isolation and identification were performed using standard and polymerase chain reaction (PCR) methods. The disc diffusion method was used to determine the resistance of Salmonella isolates to 16 antimicrobial agents and PCR to determine their serovars. Ninety-eight (64.9%) of the 151 chickens were contaminated with Salmonella of which 94.9% (93/98) were resistant serovars. The frequency of antimicrobial resistance of Salmonella isolates was high to erythromycin (94.9%) and spectinomycin (82.7%) but was low to ciprofloxacin (1.0%) and norfloxacin (1.0%) (p < 0.05). All 170 isolates of Salmonella tested exhibited resistance to one or more antimicrobial agents and the frequency varied significantly (p < 0.05) across the townships, the type of samples and the serovars. The prevalence of multidrug resistance (MDR) in Salmonella was 81.8% (139/170). Our findings pose zoonotic, food safety and therapeutic risks to workers and consumers of undercooked, contaminated chickens from these outlets.

Q Fever: Seroprevalence, Risk Factors in Slaughter Livestock and Genotypes of Coxiella burnetii in South Africa.

Q fever is a neglected zoonosis in South Africa, causing significant losses in livestock and game animals through reproductive disorders. However, there are limited studies on the extent of Coxiella burnetii infections in livestock in South Africa. Further, there is also lack of knowledge about the types of C. burnetii strains that are currently circulating in the country. Therefore, a cross-sectional, abattoir-based study was conducted to determine the seroprevalence of C. burnetii and associated risk factors, and to characterize C. burnetii strains from slaughter livestock at red meat abattoirs in Gauteng, South Africa. Of the 507 animals tested, 6.9% (95% CI: 4.9-9.5%) were positive for antibodies against C. burnetii. The seroprevalence was 9.4% (31/331) in cattle, 4.3% (3/69) in sheep, and 0.9% (1/107) in pigs. Out of the 63 tissue samples from 35 seropositive animals including material from two sheep aborted fetuses from Mangaung district (Free State province), 12.7% (8/63) tested positive by IS1111 PCR. Genotyping of the eight PCR-positive tissues from eight animals by MLVA revealed two novel genotypes, not available in Coxiella MLVA databases. It is concluded that slaughter animals pose a risk of exposing abattoir and farm workers to C. burnetii in South Africa.

Prevalence, Serovars, and Factors Associated with Salmonella Contamination of Chicken Carcasses Sold in Outlets of the Informal Market in Gauteng Province, South Africa.

ABSTRACT: Salmonella has been linked to many foodborne illnesses and epidemics in both humans and animals. This cross-sectional study determined the prevalence, serovars, and factors associated with Salmonella contamination of chickens slaughtered in informal market outlets in Gauteng Province, South Africa. A total of 151 chicken carcasses were randomly collected from 47 outlets. Standard bacteriological and molecular methods were used to isolate, identify, and determine the serovar of Salmonella isolates. The prevalence of Salmonella in carcass swabs, cloacal swabs, and carcass drips was 29.1% (44 of 151), 27.2% (41 of 151), and 43.7% (66 of 151), respectively, and the differences were statistically significant (P = 0.004). Only 5 (township locations of outlet, throughput, carcass evisceration, location of carcass for sale, and outlet sanitation) of 10 factors investigated for the contamination of carcasses by Salmonella were statistically significantly (P < 0.05) associated with the isolation of Salmonella. Of the 268 isolates of Salmonella, 157 (58.6%) were typeable using a limited molecular PCR technique, and nine serovars were identified. The predominant Salmonella enterica serovars were Bovismorbificans (31.0%), Enteritidis (7.5%), and Hadar (6.7%). The five important factors found to be significantly associated with the isolation of Salmonella at these outlets offer opportunities for the reduction of Salmonella contamination. There is a need for further investigation of the probable causes of the predominant isolation of Salmonella serovar Bovismorbificans in chickens and its potential implications for human infections in South Africa. It is concluded that chickens purchased from the informal market in Gauteng Province can be a source for salmonellosis in humans if improperly cooked before consumption.

Occurrence and Concentrations of Residues of Tetracyclines, Polyether Ionophores, and Anthelmintics in Livers of Chickens Sold in the Informal Market in Gauteng Province, South Africa.

ABSTRACT: The occurrence, concentrations, and variables associated with tetracycline, polyether ionophore, and anthelmintic residues in the livers of chickens sold in the informal market in South Africa were determined. Ultrahigh-performance liquid chromatography-tandem mass spectrometry was used to simultaneously analyze for four tetracyclines, five polyether ionophores, and six anthelmintic residues. The study determined the presence of residues in liver samples at both the limit of quantifications (LOQ) and concentrations over the maximum residue limit (MRL), i.e., noncompliant. Doxycycline (tetracycline group) was detected in 24 (24.5%) of 98 chicken livers, and 15 (15.3%) of the 98 were noncompliant. Mean ± standard deviation (SD) concentrations of 919.04 ± 1,081.30 ppb (LOQ) and 1,410.57 ± 108.89 ppb (MRL) were obtained. Maduramicin was detected in 27 (27.6%) of 98 chicken livers, and 19 (19.4%) of 98 were noncompliant. The mean ± SD for LOQ was 117.96 ± 84.56 ppb, and MRL was 153.21 ± 76.29 ppb. The concentrations of residues of doxycycline and maduramicin in chicken livers varied significantly across townships. Lasalocid was found in 31 (31.6%) of 98 samples, of which 5 (5.1%) had concentrations above the MRL. The mean ± SD concentration of lasalocid was 62.90 ± 170.84 ppb for samples in which lasalocid was quantified and 310.16 ± 356.68 ppb for noncompliant samples. Detectable concentrations of praziquantel, closantel, and rafoxanide (anthelmintics) residues were found in 3 (3.1%), 1 (1.0%), and 2 (2.0%) of 98 chicken livers, respectively. The presence of residues of three classes of veterinary drugs in chicken livers poses food safety implications to consumers and indicates a need for enhanced regulatory enforcement in controlling these drugs in South Africa.

A retrospective study (2007-2015) on brucellosis seropositivity in livestock in South Africa.

In South Africa, brucellosis testing and record-keeping are done by several laboratories, thus it is difficult to access any organized data to assess the status of the disease. This study evaluated the seropositivity for brucellosis using Rose Bengal test and complement fixation test in suspect cattle, sheep, goats and pigs sera submitted to Bacterial Serology Laboratory, Agricultural Research Council-Onderstepoort Veterinary Research (ARC-OVR) from nine provinces in the country during the period 2007-2015. This retrospective data analysis was conducted to estimate the occurrence of brucellosis in the country from the submitted samples, identify variables that affected seropositivity for brucellosis, investigate existing gaps in data recording and make recommendations on important variables to facilitate better data capture and inferences on brucellosis. Nine years of data were collated and analysed to detect association (seropositivity over time regarding animal species and location). Of the 764,276 animals tested, the distribution of samples was 90.50% (691,539/764,276), 5.19% (39,672/764,276), 3.92% (29,967/764,276) and 0.41% (3,098/764,276) for cattle, sheep, goats and pigs, respectively. The seropositivity for brucellosis by animal species was 6.31% (43,666/691,539, 95% CI: 6.26-6.37), 2.09% (828/39,672, 95% CI: 1.95-2.23), 0.63% (189/29,967, 95% CI: 0.55-0.73) and 0.13% (4/3,098, 95% CI: 0.05-0.33) in cattle, sheep, goats and pigs respectively. The data available did not capture information on the age, sex, breed and other host risk factors that would have been related to seropositivity for brucellosis. The data provide an understanding of the disease occurrence and confirm that brucellosis is enzootic in South Africa. Improved and standardized data collection can be used to pro-actively drive, monitor, change or formulate policies to mitigate the challenges brought about by brucellosis in the livestock sector in South Africa.

Occurrence of Virulence and Resistance Genes in Salmonella in Cloacae of Slaughtered Chickens and Ducks at Pluck Shops in Trinidad.

ABSTRACT: The study used PCR to determine the molecular basis of the antibiotic resistance and virulence profiles of isolates of Salmonella by targeting genes encoding for carriage and persistence in the poultry. Of a total 1,503 cecal samples collected from poultry, 91 (6.1%) were positive for Salmonella. Ten different serotypes were detected from Salmonella isolates. The study was also conducted to determine the occurrence of 13 virulence and 12 resistance genes in isolates of Salmonella. All 46 isolates of Salmonella tested were positive for one or more of the 12 virulence genes detected, ranging from 0.0% (viaB) to 100.0% (invA, mgtB, pipA, and spi4D) (P < 0.05). Occurrence of virulence genes varied significantly (P < 0.05) by serotype but not by animal species. Only 4 (33.3%) of 12 resistance genes assayed were detected: strA, ampC, cmy2, and qnrB. Overall, the occurrence of detected resistance genes was 71.7% (33 of 46), and 87.1 and 40.0% of the isolates from chickens and ducks, respectively, were positive (P = 0.0009). The occurrence of resistance genes ranged from 2.2% (cmy2) to 50.0% (qnrB) in isolates positive for resistance gene. The findings provide evidence that poultry from "pluck shops" are colonized by Salmonella pathogens that harbor virulence and antimicrobial resistance genes; this may have clinical and therapeutic consequences, if the genes detected are expressed. Although there is a need for prudent use of antimicrobial agents in poultry production systems, there should be constant monitoring for the prevalence of resistance in Salmonella isolates using phenotypic methods. The importance of monitoring the occurrence of resistance genes in the pathogens in Trinidad cannot be ignored.