Detection of Salmonella Pathogenicity Islands and Antimicrobial-Resistant Genes in Salmonella enterica Serovars Enteritidis and Typhimurium Isolated from Broiler Chickens.

Rapid growth in commercial poultry production is one of the major sources of Salmonella infections that leads to human salmonellosis. The two main Salmonella enterica serovars associated with human salmonellosis are enteritidis and typhimurium. The aim of this study was to determine the prevalence of S. enterica serovars Enteritidis and S. Typhimurium as well as their Salmonella pathogenicity islands (SPI) and antibiotic resistance profiles in broiler chicken feces from slaughterhouses. A total of 480 fecal samples from broiler chickens that were grouped into 96 pooled samples were identified to have Salmonella spp. using the invA gene, whilst the Spy and sdfI genes were used to screen for the presence of S. Enteritidis and S. Typhimurium serovars, respectively, by polymerase chain reaction (PCR) assays. The isolates were also screened for the presence of Salmonella pathogenicity islands (SPIs) using PCR. The disc diffusion assay was performed to determine the antibiotic resistance profiles of the isolates. A total of 36 isolates were confirmed as Salmonella spp. through amplification of the invA gene. Out of 36 confirmed Salmonella spp. a total of 22 isolates were classified as S. Enteritidis (n = 8) and were S. Typhimurium (n = 14) serovars. All (n = 22) S. Enteritidis and S. Typhimurium isolates possessed the hilA (SPI-1), ssrB (SPI-2) and pagC (SPI-11) pathogenicity islands genes. Amongst these serovars, 50% of the isolates (n = 11/22) were resistant to tetracycline and nalidixic acid. Only 22% of the isolates, S. Typhimurium (13.6%) and S. Enteritidis (9.1%) demonstrated resistance against three or more antibiotic classes. The most detected antibiotic resistance genes were tet(K), mcr-1, sulI and strA with 13 (59.1%), 9 (40.9%), 9 (40.9%) and 7 (31.8%), respectively. The findings of this study revealed that S. Typhimurium is the most prevalent serotype detected in chicken feces. To reduce the risk to human health posed by salmonellosis, a stringent public health and food safety policy is required.

Antibiotic resistance and virulence profiles of Proteus mirabilis isolated from broiler chickens at abattoir in South Africa.

BACKGROUND: Proteus mirabilis has been identified as an important zoonotic pathogen, causing several illnesses such as diarrhoea, keratitis and urinary tract infections. OBJECTIVE: This study assessed the prevalence of P. mirabilis in broiler chickens, its antibiotic resistance (AR) patterns, ESBL-producing P. mirabilis and the presence of virulence genes. METHODS: A total of 26 isolates were confirmed as P. mirabilis from 480 pooled broiler chicken faecal samples by polymerase chain reaction (PCR). The disk diffusion method was used to evaluate the antibacterial susceptibility test, while nine virulence genes and 26 AR genes were also screened by PCR. RESULTS: All 26 P. mirabilis isolates harboured the ireA (siderophore receptors), ptA, and zapA (proteases), ucaA, pmfA, atfA, and mrpA (fimbriae), hlyA and hpmA (haemolysins) virulence genes. The P. mirabilis isolates were resistant to ciprofloxacin (62%) and levofloxacin (54%), while 8 (30.7%) of the isolates were classified as multidrug resistant (MDR). PCR analysis identified the blaCTX-M gene (62%), blaTEM (58%) and blaCTX-M-2 (38%). Further screening for AMR genes identified mcr-1, cat1, cat2, qnrA, qnrD and mecA, 12%, 19%, 12%, 54%, 27% and 8%, respectively for P. mirabilis isolates. The prevalence of the integron integrase intI1 and intI2 genes was 43% and 4%, respectively. CONCLUSIONS: The rise of ciprofloxacin and levofloxacin resistance, as well as MDR strains, is a public health threat that points to a challenge in the treatment of infections caused by these zoonotic bacteria. Furthermore, because ESBL-producing P. mirabilis has the potential to spread to humans, the presence of blaCTX -M -producing P. mirabilis in broilers should be kept under control. This is the first study undertaken to isolate P. mirabilis from chicken faecal samples and investigate its antibiotic resistance status as well as virulence profiles in South Africa.

Indigenous knowledge and use of medicinal plants for ethnoveterinary within the North West Province, South Africa.

Ethnoveterinary medicine (EVM) has been used by local South African communities for centuries. However, the knowledge of EVM is under threat due to the influence of Western medicine and acculturation. This study aimed to document the knowledge of ethnoveterinary medicinal plants in four villages within the Mahikeng Local Municipality to help preserve this valuable knowledge. The study used a qualitative research approach and targeted practitioners and holders of ethnoveterinary knowledge in four villages. Participants were recruited using key informants and snowball techniques, and in-depth interviews were conducted using semi-structured interview schedules. The data collected was analyzed thematically, and herbarium specimens were prepared from medicinal plants during field walks and sent to the South African National Biodiversity Institute (SANBI) for identification. The study documented the use of thirty-one medicinal plant species, with Senna italica (sebetebete) emerging as the most frequently used species for multiple animal health indications. The most common diseases treated using EVM were gala (general malaise due to bile reflux) and retained placenta. Participants strongly agreed on the cultural significance of plants for treating five animal diseases, including coughs and diarrhea. Although the study revealed a high level of trust in ethnoveterinary medicine among participants, they expressed concern regarding the loss of this knowledge due to the influence of Western medicine and acculturation. The study concluded that the indigenous knowledge and use of ethnoveterinary medicine was prevalent among livestock owners in the communities of the North West Province, and more studies need to be conducted in other areas to preserve this valuable knowledge.

A Single Question on Maternal HIV Status Can Improve Identification of Children Living With HIV in South Africa.

INTRODUCTION: Screening tools to improve identification of children living with HIV (CLHIV) have been validated and used in various settings. The aim of our study was to optimize a screening tool for Primary Healthcare Clinics (PHCs) in South Africa (SA). METHODS: A cross-sectional study was conducted at PHCs in Johannesburg and Mopani Districts, between June 2021 and June 2022. Children 5-14 years of age with HIV negative or unknown status accompanied by their mothers, or appropriate caregivers, were enrolled. Demographic data, responses to the screening tool questions, and HIV test results were captured. Logistic regression modeling was used to optimize an existing 10-item screening tool, and sensitivity, specificity, and number needed to test (NNT) used to choose the final tool. RESULTS: We enrolled 14,147 children in the study, with 62 children testing HIV positive (HIV positivity of 0.4%). The 10-item tool with a single positive response had a sensitivity of 91.9% and specificity of 43.3%. An optimal combination of 5-items with two positive responses had the lowest NNT of 72, 82.3% sensitivity and 74.2% specificity. Maternal HIV status alone, HIV positive or unknown, had a 95.2% sensitivity, 65.0% specificity and NNT of 84. The 1-item tool only would have missed 5% of CLHIV (N = 3) compared with the 5-item tool that missed 18% (n = 11). CONCLUSIONS: A 1-item screening tool asking about maternal HIV status can improve efficiency of testing of children in primary healthcare facilities in SA and improve identification of CLHIV who are not on treatment.

A systematic review and meta-analysis on prevalence and antimicrobial resistance profile of Escherichia coli isolated from water in africa (2000-2021).

Water is essential for the survival of humans, animals and plants. Numerous research has been conducted on the prevalence and antibiotic resistance of Escherichia coli (E. coli) in water from various African countries, however, there is lack of comprehensive analysis of published literature. We conducted a systematic review and meta-analysis following the PRISMA guidelines where articles published in English language between January 2000 and March 2022 were searched from ScienceDirect, PubMed, Google Scholar, Scopus, African Journal Online (AJO), and Africa Index Medicus (AIM). Comprehensive Meta-Analysis (CMA) Ver 3.0 software was used to analyze the data. The pooled prevalence estimate (PPE) with 95% confidence interval was calculated using the random-effects model (CI). The overall PPE and antimicrobial resistance trends of E. coli isolated from water was screened from 4009 isolates which were isolated from 2586 samples. We extracted data from 17 studies including drinking water (n = 6), rivers (n = 5), wastewaters (n = 4) and wastewater/river (n = 1) which are all covering 27 countries in Africa with 3438 isolates. The PPE of E. coli in water was 71.7% (0.717; 95% CI: 0.562-0.833). The highest PPE antibiotic resistance was against penicillin followed by erythromycin, and ampicilin with resistance rates of 93.4%, 92.3%, and 69.4%, respectively. This systematic review provides critical evidence of E. coli consolidated prevalence and antibiotic resistance profiles, as well as regions where future studies and enhanced reporting could be beneficial in the African continent.

Prevalence and antimicrobial resistance profiles of Campylobacter species in South Africa: a "One Health" approach using systematic review and meta-analysis.

OBJECTIVES: This study investigated the prevalence and antibiotic resistance (AR) profiles of Campylobacter spp. isolated from animals, humans, and the environment in South Africa based on available published data. METHODS: Original articles published from January 1, 1990 to January 1, 2021 were searched from PubMed, ScienceDirect, Google Scholar, Africa Index Medicus, Scopus, and African Journal Online databases. Data were analyzed with Comprehensive Meta-Analysis (version 3.0). RESULTS: After screening, articles on animals (n = 25), humans (n = 7), environment (n = 3), animals/environment (n = 2), and a (n = 1) study on animals, humans, and the environment were included in this review. The pooled prevalence estimates (PPEs) were 28.8%, 16.4%, and 28.4% in animals, humans, and the environment, respectively. The Campylobacter jejuni and Campylobacter coli species were commonly isolated from humans, animals, and the environment in South Africa. The AR profiles were screened from 2032 Campylobacter spp., with the highest PPE of AR observed against clindamycin (76.9%) and clarithromycin (76.5%). Campylobacter isolates tested with the disk diffusion assay and minimum inhibitory concentration methods recorded an overall AR prevalence of 35.3% and 37.1%, respectively, whereas multidrug resistance PPE was 35.3%. CONCLUSION: Regular surveillance of Campylobacter spp. prevalence and its antimicrobial resistance strains is recommended, as well as the formulation of a "One Health" approach for better management and control of Campylobacter spp. infection in South Africa.

Campylobacter jejuni from Slaughter Age Broiler Chickens: Genetic Characterization, Virulence, and Antimicrobial Resistance Genes.

Campylobacter jejuni is a major cause of food-borne human gastroenteritis worldwide and is designated as a high priority antimicrobial-resistant pathogen by the World Health Organization (WHO). In this study, a total of 26 C. jejuni isolates from broiler chickens were screened for the presence of virulence and antimicrobial resistance genes by PCR. As a result, the study detected 11/26 (42.3%), 9/26 (34.6%), 8/26 (30.8%), 7/26 (26.9%), 6/26 (23.1%), and 6/26 (23.1%) of cdtC, pldA, cdtB, cdtA, cadF, and ciaB virulence genes, respectively, with seven of the isolates carrying more than two virulence genes. The majority of the isolates n = 25 (96.1%) were resistant to nalidixic acid, followed by n = 21 (80.7%), n = 22 (84.6%), and n = 5 (19.2%) for tetracycline, erythromycin, and ciprofloxacin, respectively. Most isolates were harboring catI (n = 16; 84.2%), catII (n = 15; 78.9%), catIII (n = 10; 52.6%), catIV (n = 2; 10.5%), floR (n = 10; 52.6%), ermB (n = 14; 73.7%), tetO (n = 13; 68.4%), tetA (n = 9; 47.4%), mcr-4 (n = 8; 42.1%), and ampC (n = 2; 10.5%). Meanwhile, mcr-1, mcr-2, mcr-3, mcr-5, tet(X), tet(P), and tet(W) genes were not detected in all isolates. Class I and Class II integrons were detected in 92.3% (n = 24) and 65.4% (n = 17) isolates, respectively. About 31% (8 of the 26 isolates) isolates were carrying more than two resistance genes. According to our knowledge, this is the first study to detect class II integrons in Campylobacter spp. (C. jejuni). The high prevalence of cdtA, cdtB, cdtC, cadF, pldA, and ciaB genes and antibiotic resistance genes in C. jejuni in this study indicates the pathogenic potential of these isolates. Majority of the isolates demonstrated resistance to nalidixic acid, tetracycline (tet), and erythromycin (ermB), which are the drugs of choice for treating Campylobacter infections. Therefore, these findings highlight the importance of implementing an efficient strategy to control Campylobacter in chickens and to reduce antimicrobial use in the poultry industry, which will help to prevent the spread of infections to humans.

Outcomes of a Telephonic Postnatal Intervention for Mothers and Babies in Mopani District, Limpopo, South Africa.

Background: The postnatal period is a critical period for the health of both mother and infant. Studies show that postnatal care reduces neonatal mortality and other adverse mother and child health outcomes. While the World Health Organization recommends four postnatal care contacts, South African guidelines only specify three, excluding a 7-14-day post-birth contact. This study aimed to assess whether a telephonic contact at 7-14 days following delivery had any effect on use of additional postnatal services. Methods: A randomized controlled trial design was used to address the study objectives. Two groups of new mothers were randomly allocated to either receive the 7-14-day telephonic contact or not from a research nurse. Data for this study was collected at Maphutha L Malatjie Hospital (MLMH). Descriptive analysis was performed first, then a multivariable logistic regression analysis was conducted to assess the factors associated with access to other health care services. Results: A total of 882 mothers were recruited, 854 (97%) were classified as high risk, 28 (3%) were classified as low risk. 417 (49%) of the high risk received the 7-14-day call (intervention group) whilst the remainder of 437 (51%) from the high risk plus all mothers classified as low risk (28) did not receive the call (control group). 686 (78%) of all mothers received the 3 month follow up call. The call showed that 17 mothers from the control group and 10 mothers from the intervention group accessed other healthcare services. We find that hypertension (3.28; 1.06 -10.10), mental health risk (2.82; 1.25 -6.38), PV bleeding during pregnancy (18.33; 1.79-187.61), problem during labor (4.40; 1.280-15.13) were positively associated with access to other health services, with statistically significant associations (p-value < 0.05). We found statistically insignificant associations between receiving the 7-14-day call and accessing other health care services. Conclusion: The 7-14-day call had no statistically significant impact on access to other health services, however, high levels of satisfaction with the call may point to an unmet need for care at this time. It is important to investigate other innovative solutions to postnatal care improvement in South Africa.

Molecular Detection of Integrons, Colistin and ß-lactamase Resistant Genes in Salmonella enterica Serovars Enteritidis and Typhimurium Isolated from Chickens and Rats Inhabiting Poultry Farms.

The rapid growth of multidrug-resistant Salmonella is a global public health concern. The aim of this study was to detect integrons, colistin and ß-lactamase resistance genes in Salmonella enteritidis and typhimurium. A total of 63 isolates of S. enteritidis (n = 18) and S. typhimurium (n = 45) from fecal samples of layers and rats at chicken farms were screened for antibiotic resistant genes. Conventional PCR was performed for the detection of integrons (classes 1, 2, and 3), colistin (mcr-1-5) and ß-lactamase (blaCTX-M, blaCTX-M-1, blaCTX-M-2, blaCTX-M-9, blaCTX-M-15, blaTEM, blaSHV, and blaOXA) resistant genes. Of these isolates, 77% and 27% of S. typhimurium and S. enteritidis harboured the mcr-4 encoded gene for colistin, respectively. The prevalence of class 1 integrons for S. typhimurium and S. enteritidis was 100% for each serovar, while for class 2 integrons of S. typhimurium and S. enteritidis it was 49% and 33% respectively, while class 3 integron genes was not detected. Our study also detected high levels of ß-lactamase encoding genes (bla gene), namely blaCTX-M, blaCTX-M-1, blaCTX-M-9 and blaTEM from both S. typhimurium and S. enteritidis. This, to our knowledge, is the first report of mcr-4 resistance gene detection in Salmonella serovars in South Africa. This study also highlights the importance of controlling rats at poultry farms in order to reduce the risk of transmission of antibiotic resistance to chickens and eventually to humans.

Risk profile of postnatal women and their babies attending a rural district hospital in South Africa.

Background: Maternal and neonatal mortality remain unacceptably high and inequitably distributed in South Africa, with the postnatal period being a dangerous time for both mother and baby. The aim of this paper is to describe the risk factors for poor postnatal outcomes, including postnatal mental health disorders, in a population of postnatal women and their babies utilising rural district hospital services in Limpopo Province, with a focus on HIV. We also describe health care provider compliance with relevant guidelines. Methods: All women discharged from the postnatal ward of the district hospital who consented to participate were enrolled. A research nurse used a structured questionnaire to collect data about sociodemographic information, pregnancy and pre-existing conditions, complications during labour and birth, pregnancy outcomes and mental health risk factors. Results: The questionnaire was completed for 882 women at the time of discharge. Only 354 (40.2%) of participants had completed secondary education, and 105 (11.9%) reported formal employment. Chronic hypertension was recorded in 20 women (2.3%), with an additional 49 (5.6%) developing a hypertensive disorder during pregnancy. HIV prevalence was 22.8%. 216 women (24.5%) had a mental health risk factor, with 40 reporting more than one (4.5%). Having no income, no antenatal care, having HIV and any hypertensive disorder were significantly associated with a positive mental health risk screen in multivariable analysis. There were 31 stillbirths and early neonatal deaths (3.5%), and 119 babies (13.4%) were born at a low birth weight. Stillbirth or early neonatal death was significantly associated with no antenatal care in multivariable analysis. Conclusions: Women and babies in this study experienced multiple risk factors for poor outcomes in the postpartum period. Postnatal care should be strengthened in order to address the dominant risks to mothers and babies, including socioeconomic challenges, HIV and hypertension, and risks to mental health. Tools to identify mothers and babies at risk of postnatal complications would allow limited resources to be allocated where they are most needed.

A Review of Ethnoveterinary Knowledge, Biological Activities and Secondary Metabolites of Medicinal Woody Plants Used for Managing Animal Health in South Africa.

Globally, the use of ethnoveterinary medicine as remedies for animal health among different ethnic groups justify the need for a systematic exploration to enhance their potential. In addition, the increasing popularity and utilisation of woody plants remain common in traditional medicine, which may be attributed to their inherent benefits. The current review was aimed at analysing ethnoveterinary surveys, biological activities, and secondary metabolites/phytochemical profiles of the woody plants of South Africa. Eligible literature (period: 2000 to 2020) were retrieved from different databases such as Google Scholar, PubMed, Sabinet, and Science Direct. Based on the inclusion and exclusion criteria, 20 ethnoveterinary surveys were eligible and were subjected to further analysis. We identified 104 woody plant species from 44 plant families that are used in the treatment of different diseases in animals, particularly cattle (70%) and goats (20%). The most mentioned (with six citations) woody plants were Terminalia sericea Burch. ex DC and Ziziphus mucronata Willd., which were followed by plants with five (Cussonia spicata Thunb., Pterocarpus angolensis DC and Vachellia karroo (Hayne) Banfi & Galasso) or four (Acokanthera oppositifolia (Lam.) Codd, Cassia abbreviata Oliv., and Strychnos henningsii Gilg) individual mentions. The most dominant families were Fabaceae (19%), Apocynaceae (5.8%), Rubiaceae (5.8%), Anacardiaceae (4.8%), Combretaceae (4.8%), Euphorbiaceae (4.8%), Malvaceae (4.8%), Rhamnaceae (4.8%), and Celastraceae (3.8%). Bark (33%), leaves (29%), and roots (19%) were the plant parts dominantly used to prepare remedies for ethnoveterinary medicine. An estimated 20% of woody plants have been screened for antimicrobial, anthelmintic, antioxidant, and cytotoxicity effects. Phytochemical profiles established a rich pool of valuable secondary metabolites (phenolic, flavonoids and condensed tannins) that may be responsible for the exerted biological activities. Overall, the significant portion of woody plants lacking empirical evidence on their biological effects indicates a major knowledge gap that requires more research efforts.

Isolation and antibiotic sensitivity of Campylobacter species from fecal samples of broiler chickens in North West Province, South Africa.

BACKGROUND AND AIM: Infections with Campylobacter species have gained recognition as the most frequent cause of foodborne gastroenteritis globally. Their significance in South Africa is still an area of study interest. This study was, therefore, carried out to determine the occurrence of Campylobacter species in chickens from North West Province of South Africa as well as their antibiotic sensitivity status. MATERIALS AND METHODS: A total of 2400 chicken fecal samples were collected and pooled to a total of 480 samples from five registered active poultry abattoirs in the Ngaka Modiri Molema District of North West Province, South Africa. Polymerase chain reaction (PCR) was used for the detection of Campylobacter spp. targeting the 16S rRNA gene while antibiotic sensitivity was determined using disk diffusion inhibition test. RESULTS: After isolation, a total of 26 samples were confirmed to be harboring Campylobacter jejuni by PCR and sequencing. C. jejuni was found to be the only isolate detected in all the fecal samples tested. The study further demonstrated that C. jejuni infections were highest in the summer season (3%) followed by autumn and winter at 1%, while there were none detected in the spring. The isolated C. jejuni-positive samples on disk diffusion inhibition test displayed resistance to nalidixic acid, tetracycline, erythromycin, and ciprofloxacin at 98%, 80%, 83%, and 21%, respectively. CONCLUSION: C. jejuni isolated in this study is known to cause disease in humans, and thus its occurrence requires application of "One Health" strategy to reduce the spread of this zoonotic pathogen in South Africa.

Canine aflatoxicosis outbreak in South Africa (2011): a possible multi-mycotoxins aetiology.

This study analysed 60 dog food samples obtained from commercial outlets following the 2011 aflatoxicosis outbreak in South Africa. Results obtained from the selected dog food samples revealed that 87% of samples were contaminated with aflatoxins (AFs) (mainly AFB1 and AFB2). Amongst these samples, 45 (75%) were above the 20 parts per billion (ppb) set by most countries and 10 ppb regulated by the Fertilizers, Farm Feeds, Agricultural Remedies and Stock Remedies Act (Act No. 36 of 1947) for South Africa. In addition to AFs, other mycotoxins were also detected in the same samples with fumonisins (FBs) (mainly FB1 and FB2) contaminating 98% of samples with 49 (81.81) above the tolerable limit of 1000 ppb in feedstuff set up by the Federal Drug Agency (FDA) (USA). The FBs mean obtained was 1556 ppb (Table 1) with contamination varying between 5.2 and 4653.8 ppb. Ochratoxin A (OTA) was detected in 41 (68%) of the analysed samples, with a mean value of 13.7 ppb. Amongst these samples, 15 (25%) were above the 20 ppb highest limit set by the Codex Alimentarius standard. Zearalenone (ZEA) was detected in 96% of samples, with a mean value of 354.1 ppb. Thirty-three samples (55%) were above the regulated level 1000 ppb tolerable limit. The recoveries were up to ten times the tolerable daily limits of the FDA and EU. The correlation between mycotoxin findings and clinical signs reported on patients presented for aflatoxicosis led to the conclusion that the outbreak was associated with the presence of other mycotoxins found simultaneously in the analysed samples by additive or synergistic effects.