Non-typhoidal Salmonella contributes to gastrointestinal infections in Morogoro: Evidence from patients attending Morogoro regional referral hospital in Tanzania.

INTRODUCTION: Salmonella is one of the most common causes of food-borne outbreaks and infection worldwide. Non-typhoidal Salmonella (NTS) infections are common and remain a significant public health problem among important bacterial foodborne diseases. The current study aimed to establish the Non typhoidal Salmonella infection and antimicrobial resistance status among selected patients at Morogoro Regional Referral Hospital (MRRH), Morogoro Region, Tanzania, to inform clinical care management and public health interventions. METHODOLOGY: A cross-sectional study was conducted using medical records and samples were collected from hospitalised and outpatients between October and December 2021. A total of 153 participants were enrolled in the study and 132 consented to being sampled. The collected samples were analysed using standard microbiological techniques. The isolates were subjected to molecular genotyping, where Polymerase Chain Reaction (PCR) was performed targeting the 16S rDNA gene. PCR products were then submitted for sequencing to establish phylogenetic relatedness. Antimicrobial susceptibility testing and resistance genes screening were also conducted. RESULTS: The phylogenetic analysis identified two Salmonella serovars; Salmonella Enteritidis and Salmonella Typhimurium. The isolates were from four adults and seven children patients. The isolates were tested against six antimicrobial agents: tetracycline, trimethoprim, gentamycin, ciprofloxacin, ampicillin and cefotaxime. Further antimicrobial assays were performed by screening 10 antimicrobial resistance genes using PCR. Overall, the highest resistance was observed in ampicillin (100%), whereas the lowest resistance was recorded for ciprofloxacin and gentamicin (9.1%). In addition, four (36.4%) of the isolates were resistant to cefotaxime and three (27.3%) to tetracycline and trimethoprim. The isolates also exhibit the presence of resistance genes for sulfamethoxazole 1&2, tetracycline (tet) A&B, Beta-lactamase CTXM, Beta-lactamase TEM, Beta-lactamase SHV, Gentamycine, Acra and acc3-1 in different occurrences. The overall prevalence of Salmonella species in Morogoro region was 8.3% (11/132) with Salmonella Enteritidis and Salmonella Typhimurium being the only serovars detected from adults and children stool samples. CONCLUSION: Our investigation showed that both children and adults had been exposed to Salmonella spp. However, the occurrence of NTS was higher in children (5.3% (7/132) compared to adults (3.0% (4/132). To stop zoonotic infections and the development of antimicrobial resistance in the community, this calls for Infection Prevention and Control (IPC) and stewardship programmes on rational use of antimicrobials in both health facilities and at the community level.

Seroprevalence and risk factors for Taenia spp infection in pigs in Kongwa and Songwe districts, Tanzania: A cross-sectional study.

Taenia solium porcine cysticercosis (PCC) is widespread in many low- and middle-income countries (LMICs) where free-range pig rearing is common and hygienic standards are subpar. A cross-sectional survey was conducted in 42 villages between June and September 2019 (14 in Songwe district, southwest Tanzania, and 28 in Kongwa district, central Tanzania). Using a commercial Ag-ELISA kit (apDia, Belgium), circulating antigens of Taenia spp in pig serum were identified and used to calculate the PCC seroprevalence. The study recruited 692 randomly selected households, sampling one pig per household. The relationship between each risk factor and the seroprevalence of PCC at the household and village levels was analysed using mixed logistic regression models. The findings showed that approximately 28% of the pigs were reared in free-range settings, the proportion of households with latrines across the districts was 92%. Twenty-seven percent of households with latrines had water and soap available for hand washing. Sixty-seven (9.7%) tested positive for PCC based on Ag-ELISA. The overall seroprevalence in Kongwa and Songwe districts was 7.3% and 14.0% respectively. In addition, the overall village Ag-ELISA positivity was 9.3%, with an interquartile range (IQR) of 4.6% - 14.1%. Increasing the age of the pig (OR = 3.13 95% CI = 1.48 - 6.60; p = 0.003), pig originating from outside the household (OR = 0.5 95% CI = 0.25 - 0.99; p = 0.05), and pigs kept in a household that practised deworming (OR = 2.23 95% CI = 1.08 - 4.61; p = 0.03) were important risk factors associated with PCC positivity. Therefore, the high seroprevalence of PCC, up to 14%, calls for rapid and effective control actions such as vaccination and treatment of pigs against PCC, and public health education emphasises on indoor pig rearing, hygienic practices and regular use of latrines. Our findings also point to a potential danger of Taenia. spp infection indicating the possibility of people carrying the adult parasite Taenia solium not only in the rural communities of Kongwa and Songwe districts but also in the urban areas of Tanzania, where pigs from these areas are transported for consumption. To develop effective management measures, further research on taeniasis and cysticercosis in the human population is required.

Changes in fatty acids during storage of artisanal-processed freshwater sardines (Rastrineobola argentea).

For ages, indigenous small fish species have been important in food and nutritional security of poor communities in low income countries. Freshwater fish, in particular fatty fish species are attracting a great attention because they are good sources of health promoting long chain omega-3 fatty acids. Docosahexaenoic acid (DHA, C22:6n-3), Docosapentaenoic acid (DPA, C22:5n-3) and eicosapentaenoic acid (EPA, C20:5n-3) are the main omega-3 PUFAs known to confer health benefits in humans if consumed in required amounts. While nutritionally valued, omega-3 PUFAs in fish are susceptible to oxidative damage during processing, transportation and subsequent storage. Lake Victoria sardines (Rastrineobola argentea), are rich source of chemically unstable omega-3 fatty acids DHA, DPA and EPA. Traditionally, sardines are preserved by sun drying, deep frying and smoking. Sardine products are transported, stored and marketed at ambient temperatures. Generally, uncontrolled and higher temperatures are known to increase vulnerability of polyunsaturated fatty acids to oxidation which in turn results into loss of nutritional and sensory qualities. This study investigated changes of fat acids in sun dried, deep fried and smoked sardines during storage. Lipolysis and the progressive hydroperoxides formation were monitored by free fatty acids (FFAs) and peroxide value (PV) respectively. None volatile secondary products of lipid oxidation were measured by thiobabituric acid reactive substances (TBARS). Fatty acids were analyzed by gas chromatography with a flameionization detector (GC-FID). Deep fried sardines maintained the lowest and apparently stable PV, TBARS and FFAs. Proportions of saturated fatty acids and polyunsaturated fatty acids decreased with time while that of monounsaturated fatty acids increased. Omega-3 fatty acids EPA, DPA and DHA decreased with increase in storage time. In 21 days of storage, DHA was oxidized beyond detectable levels in all sardine products. Gradual increase in FFAs in sun dried sardines was suggestive of lipid hydrolysis induced by enzymes.

Mitigating antimicrobial resistance (AMR) using implementation research: a development funder's approach.

Despite the escalating burden of antimicrobial resistance (AMR), the global response has not sufficiently matched the scale and scope of the issue, especially in low- and middle-income countries (LMICs). While many countries have adopted national action plans to combat AMR, their implementation has lagged due to resource constraints, dysfunctional multisectoral coordination mechanisms and, importantly, an under-recognized lack of technical capacity to adapt evidence-based AMR mitigation interventions to local contexts. AMR interventions should be tailored, context-specific, cost-effective and sustainable. The implementation and subsequent scale-up of these interventions require multidisciplinary intervention-implementation research (IIR). IIR involves both quantitative and qualitative approaches, occurs across a three-phase continuum (proof of concept, proof of implementation and informing scale-up), and across four context domains (inner setting, outer setting, stakeholders and the implementation process). We describe the theoretical underpinnings of implementation research (IR), its various components, and how to construct different IR strategies to facilitate sustainable uptake of AMR interventions. Additionally, we provide real-world examples of AMR strategies and interventions to demonstrate these principles in practice. IR provides a practical framework to implement evidence-based and sustainable AMR mitigation interventions.

Optimization of HPLC-MS/MS method for determination of antimalarial adulterants in herbal products.

The use of herbal products is booming all over the world because of being believed as safer than conventional drugs and free of side effects. However, there are untrustworthy manufacturers who adulterate herbal products by adding conventional drugs which might eventually lead to microbial resistance and herb-to-drug interactions. There is a need to develop methods for detecting adulterants in herbal products. A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneous identification and determination of conventional antimalarials (chloroquine, quinine, sulfadoxine, pyrimethamine, mefloquine, lumefantrine, amodiaquine, artemisinin, dihydroartemisinin, artesunate and artemether) in herbal products was developed. Stable isotopically labelled compounds (artemether-d3, quindine-d3, and sulfadoxine-d3) were used as internal standards (ISs) for quantitative analysis. Extraction of analytes was performed using methanol: water: formic acid (90:10:0.1, v/v) and chromatographic separation was done in a gradient mode using mobile phase A: Ultrapure water containing 0.1% formic acid and 1 mM ammonium formate and mobile phase B: Acetonitrile/methanol (50:50) containing 0.1% formic acid and 1 mM ammonium formate. The calibration curves were linear (r2 ≥ 0.991) over the range of 0.001-0.3 µg mL-1 for all compounds. The limit of detection (LOD) ranged from 0.002 to 0.02 µg mL-1 while the limit of quantification (LOQ) ranged from 0.006 to 0.08 µg mL-1. Accuracy, expressed as recovery of spiked herbal products ranged from 52 to 128%. The precision, expressed as percent relative standard deviation (%RSD) at two concentration levels, ranged from 1.0 to 13.8%. The matrix effect expressed as the matrix factor (MF) ranged from 0.77 to 0.97. The developed method was used to identify and quantify conventional antimalarials in herbal product samples from Tanzania. Ten out of 50 herbal products were found to contain amodiaquine, sulfadoxine, pyrimethamine, mefloquine, dihydroartemisinin, artemether and lumefantrine. The developed method is considered a valuable tool for getting a better understanding of the adulteration of conventional antimalarials in herbal products.

A HPLC-MS/MS method for screening of selected antibiotic adulterants in herbal drugs.

The use of herbal products adulterated with conventional drugs increases the risk of developing microbial resistance and causes herb-to-drug interaction, leading to severe clinical consequences. The complex nature of herbal products has been a challenge for the unambiguous identification of adulterants. The improved analytical selectivity and sensitivity of hyphenated techniques such as high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) enable the confirmatory screening of adulterants in herbal products. Simultaneous screening of adulterants is necessary and efficient because it has been established that more than one chemical adulterant may be present in one herbal product. An HPLC-MS/MS method for the simultaneous detection and quantification of amoxicillin, ampicillin, metronidazole, ciprofloxacin, sulfamethoxazole, and trimethoprim in powdered herbal drugs was developed. Deuterated metronidazole-d3, trimethoprim-d3, ciprofloxacin-d8, and sulfamethoxazole-d4 were used as internal standards (ISs). For each analyte, two transitions were monitored using protonated molecules as precursor ions. The extraction of analytes from herbal products was performed using a simple methanol : water : formic acid (90 : 10 : 0.05, v/v) extraction solvent. Chromatographic separation was done in a gradient of 0.01% formic acid in methanol and 0.01% formic acid in MilliQ water. The calibration curves were linear (r2 ≥ 0.996) over the range of 0.005-2.5 µg mL-1 for all compounds except metronidazole, whose range was 0.005-1 µg mL-1. The limit of detection (LOD) ranged from 0.012 to 0.046 µg mL-1, while the limit of quantification (LOQ) ranged from 0.066 to 0.153 µg mL-1. The accuracy, expressed as the recovery of spiked herbal products, ranged from 45% to 114%. The precision, expressed as relative standard deviation (RSD) at two concentration levels, ranged from 1.6% to 15.9%. The matrix effect expressed as the matrix factor (MF) ranged from 0.79 to 0.92. The developed method was applied to powder herbal products purchased in Tanzania. Amoxicillin, ampicillin, trimethoprim, sulfamethoxazole, and ciprofloxacin were not detected in all samples. Metronidazole was detected in eight samples with the highest concentration of 1.38 µg g-1. The developed method is suitable for the detection of all the studied antibiotic adulterants in herbal products. Quantification can be performed for all the compounds except ciprofloxacin due to its lower recovery.

Antimicrobial Use, Residues, Resistance and Governance in the Food and Agriculture Sectors, Tanzania.

All infections are potentially curable as long as the etiological agents are susceptible to antimicrobials. The increased rate at which antimicrobials are becoming ineffective is a global health risk of increasing concern that threatens withdrawal of beneficial antimicrobials for disease control. The increased demand for food of animal origin, in particular eggs, meat and milk has led to intensification and commercial production systems where excessive use and misuse of antimicrobials may prevail. Antimicrobials, handled and used by farmers and animal attendants with no formal education, may be predisposed to incorrect dosages, misuse, incorrect applications and non-adherence to withdrawal periods. This study was conducted to assess the regulatory roles and governance of antimicrobials, establish the pattern and extent of their use, evaluate the antimicrobial residues and resistance in the food animals and crop agriculture value chains, and relate these findings to existing strategies in place for combating the emergence of antimicrobial resistance in Tanzania. A multimethod approach (desk review, field study and interviews) was used. Relevant establishments were also visited. High levels of resistance to penicillin G, chloramphenicol, streptomycin and oxytetracycline have been reported, especially for Actinobacter pyogenes, Staphylococcus hyicus, Staphylococcus intermedius and Staphylococcus aureus from dairy cattle with mastitis and in humans. Similar trends were found in poultry where eggs and meat are contaminated with Escherichia coli strains resistant to amoxicillin + clavulanate, sulphamethoxazole and neomycin. An increasing trend of emerging multidrug resistant E. coli, Klebsiella pneumoniae, Staphylococcus aureus and Salmonella was also found in food animals. An increase in methicillin resistant Staphlococcus aureus (MRSA) and extended-spectrum beta-lactamase (ESBL) in the livestock sector in Tanzania have been reported. The pathogens isolated in animals were resistant to ampicillin, augmentin, gentamicin, co-trimoxazole, tetracycline, amoxicillin, streptomycin, nalidixic acid, azithromycin, chloramphenicol, tylosin, erythromycin, cefuroxime, norfloxacin and ciprofloxacin. An increased usage of antimicrobials for prophylaxis, and therapeutics against pathogens and for growth promotion in livestock, aquaculture and crop production were observed. A One Health strategic approach is advocated to combat antimicrobial resistance (AMR) in the food and agriculture sectors in Tanzania. Practical recommendations include (a) legislation review and implementation; (b) antimicrobial use (AMU), AMR and antimicrobial residue (AR) awareness and advocacy among stakeholders along the value chain; (c) strengthening of surveillance and monitoring programs for AMU, AMR and AR; (d) enhanced development and use of rapid and innovative diagnostic tests and the promotion of biosecurity principles; and (e) good husbandry practices. The utilization of this information to improve public health policies and reduce the burden of AMR will be beneficial.

Where and when to vaccinate? Interdisciplinary design and evaluation of the 2018 Tanzanian anti-rabies campaign.

OBJECTIVES: Hoping to improve health-related effectiveness, a two-phase vaccination against rabies was designed and executed in northern Tanzania in 2018, which included geo-epidemiological and economic perspectives. METHODS: Considering the local bio-geography and attempting to rapidly establish a protective ring around a city at risk, the first phase intervened on sites surrounding that city, where the population density was lower than in the city at risk. The second phase vaccinated a rural area. RESULTS: No rabies-related case has been reported in the vaccinated areas for over a year post-immunisation; hence, the campaign is viewed as highly cost-effective. Other metrics included: rapid implementation (concluded in half the time spent on other campaigns) and the estimated cost per protected life, which was 3.28 times lower than in similar vaccinations. CONCLUSIONS: The adopted design emphasised local bio-geographical dynamics: it prevented the occurrence of an epidemic in a city with a higher demographic density than its surrounding area and it also achieved greater effectiveness than average interventions. These interdisciplinary, policy-oriented experiences have broad and immediate applications in settings of limited and/or time-sensitive (expertise, personnel, and time available to intervene) resources and conditions.

Genome-Based Analysis of Extended-Spectrum ß-Lactamase-Producing Escherichia coli in the Aquatic Environment and Nile Perch (Lates niloticus) of Lake Victoria, Tanzania.

Extended-spectrum ß-lactamase (ESBL)-producing bacteria constitute an emerging global health issue with food products being vehicles of transmission and the aquatic environments serving as potential reservoirs. This study aimed to characterize ESBL-producing Escherichia coli in Nile perch and water from Lake Victoria in Tanzania. A total of 180 samples of Nile perch and 60 water samples were screened for ESBL-producing E. coli on MacConkey agar supplemented with 2 µg/ml of cefotaxime and confirmed by bla CTX-M and bla TEM PCR. Antimicrobial resistance was determined by the disk diffusion method, and the ESBL-producing isolates were whole genome sequencing (WGS). ESBL-producing E. coli were detected in eight of the 180 analyzed Nile perch samples, and only one water sample was positive (1.7%, n = 60). Isolates were resistant to sulfamethoxazole-trimethoprim (100%), ampicillin/cloxacillin (100%), erythromycin 72.7% (8/11), tetracycline 90.9% (10/11), and nalidixic acid 63.6% (7/11). This mostly corroborates the resistance genes that they carried for sulfonamides (sul1 and sul2), trimethoprim (dfrA and dfrB), aminoglycosides [aac(3)-IId, strA, and strB], tetracycline [tet(B) and tet(D)], and fluoroquinolones (qepA4). They harbored plasmid replicon types IncF, IncX, IncQ, and Col and carried bla CTX-M- 15 and bla TEM- 1 B genes generally found on the same contigs as the IncF plasmid replicon. Although epidemiologically unrelated, the strains formed three separate sequence type-phylogroup-serotype-specific clusters: C1, C2, and C3. Cluster C1 included five strains (3 to 13 SNPs) belonging to ST167, phylogroup A, and serotype O9:H21; the two C2 strains (11 SNPs) belong to ST156, phylogroup B1, and serotype ONT:H28; and C3 was made up of four strains (SNPs ranged from 4 to 17) of ST636, phylogroup B2, and serotype O45:H7. The common virulence gene gad was reported in all strains. In addition, strains in C2 and C3 possessed iss, lpfA, and nfaE virulence genes, and the vat gene was found only in C3. The present study reports the occurrence of multidrug-resistant ESBL-producing E. coli carrying plasmid-mediated ESBL genes in offshore water and Nile perch in Lake Victoria. Strains formed three clonal clusters of unknown origin. This study reveals that the Lake may serve as reservoir for ESBL-producing bacteria that can be transmitted by fish as a food chain hazard of One-Health concern.

Genomic insights into Vibrio cholerae O1 responsible for cholera epidemics in Tanzania between 1993 and 2017.

BACKGROUND: Tanzania is one of seven countries with the highest disease burden caused by cholera in Africa. We studied the evolution of Vibrio cholerae O1 isolated in Tanzania during the past three decades. METHODOLOGY/PRINCIPAL FINDINGS: Genome-wide analysis was performed to characterize V. cholerae O1 responsible for the Tanzanian 2015-2017 outbreak along with strains causing outbreaks in the country for the past three decades. The genomes were further analyzed in a global context of 590 strains of the seventh cholera pandemic (7PET), as well as environmental isolates from Lake Victoria. All Tanzanian cholera outbreaks were caused by the 7PET lineage. The T5 sub-lineage (ctxB3) dominated outbreaks until 1997, followed by the T10 atypical El Tor (ctxB1) up to 2015, which were replaced by the T13 atypical El Tor of the current third wave (ctxB7) causing most cholera outbreaks until 2017 with T13 being phylogenetically related to strains from East African countries, Yemen and Lake Victoria. The strains were less drug resistant with approximate 10-kb deletions found in the SXT element, which encodes resistance to sulfamethoxazole and trimethoprim. Nucleotide deletions were observed in the CTX prophage of some strains, which warrants further virulence studies. Outbreak strains share 90% of core genes with V. cholerae O1 from Lake Victoria with as low as three SNPs difference and a significantly similar accessory genome, composed of genomic islands namely the CTX prophage, Vibrio Pathogenicity Islands; toxin co-regulated pilus biosynthesis proteins and the SXT-ICE element. CONCLUSION/SIGNIFICANCE: Characterization of V. cholerae O1 from Tanzania reveals genetic diversity of the 7PET lineage composed of T5, T10 and T13 sub-lineages with introductions of new sequence types from neighboring countries. The presence of these sub-lineages in environmental isolates suggests that the African Great Lakes may serve as aquatic reservoirs for survival of V. cholerae O1 favoring continuous human exposure.

Levels and distribution of progesterone in receiving waters and wastewaters of a growing urban area.

This study aimed at investigating the levels and distribution of progesterone in receiving waters and wastewaters in Arusha, a fast-growing urban area and the third largest city in Tanzania. Specifically, the study was conducted along the Themi River and the adjacent waste stabilization ponds (WSPs). Progesterone was detected and quantified by using an enzyme-linked immunosorbent assay (ELISA) kit. For Themi River samples, the level of progesterone obtained ranged from 'no detection' to 439 ng/L with a mean value of 120.3 ng/L. The levels detected were significantly higher in the midstream than the upstream and downstream sections of the river (P<0.05). The higher values at the midstream were attributable to livestock, WSPs and household effluents; agricultural activities; and sewage infiltration. Progesterone levels in the present study, although not extremely high, correspond to those associated with harmful effects in other studies. Results for samples collected from the WSPs indicated a release of 215 ng progesterone per L of the receiving waters. Although progesterone removal efficiency by the WSPs was 75%, the amount released was still high enough to cause harm to aquatic organisms. Thus, more efficient techniques are required to adequately remove progesterone to the recommended levels.

Cholera hotspots and surveillance constraints contributing to recurrent epidemics in Tanzania.

OBJECTIVE: We described the dynamics of cholera in Tanzania between 2007 and 2017 and assessed the weaknesses of the current surveillance system in providing necessary data in achieving the global roadmap to 2030 for cholera control. RESULTS: The Poisson-based spatial scan identified cholera hotspots in mainland Tanzania. A zero-inflated Poisson regression investigated the relationship between the incidence of cholera and available demographic, socio-economic and climatic exposure variables. Four cholera hotspots were detected covering 17 regions, home to 28 million people, including the central regions and those surrounding the Lakes Victoria, Tanganyika and Nyaza. The risk of experiencing cholera in these regions was up to 2.9 times higher than elsewhere in the country. Regression analyses revealed that every 100 km of water perimeter in a region increased the cholera incidence by 1.5%. Due to the compilation of surveillance data at regional level rather than at district, we were unable to reliably identify any other significant risk factors and specific hotspots. Cholera high-risk populations in Tanzania include those living near lakes and central regions. Successful surveillance require disaggregated data available weekly and at district levels in order to serve as data for action to support the roadmap for cholera control.

Phenotypic and molecular detection of Aeromonads infection in farmed Nile tilapia in Southern highland and Northern Tanzania.

Aeromonads disease outbreaks are now becoming a common phenomenon in freshwater farmed fish worldwide. In Tanzania, the aquaculture field is increasingly growing save to sustain food protein demand and strengthen household income. To avoid losses that tilapia fish farmers might account, information on magnitude of infection and characteristics of the aetiological agent is vital. This study aimed to establish the prevalence of aeromonads infection in farmed tilapia and assess pond and fish health management practices. A cross sectional study was carried out between February 2017 and October 2018 and a total of 816 whole fish samples were aseptically collected from 32 ponds in Ruvuma, Mbeya, Iringa and Kilimanjaro regions. During sampling, water quality parameters were taken and questionnaires to assess the knowledge of farmers were also provided. Isolation and identification of bacteria was conducted using conventional biotyping and molecular techniques. A total of 201 (80.4%) of 250 isolates that were conventionally identified were confirmed to be aeromonads by amplification of 820 bp rpoD gene, making the overall prevalence of 24.6% (201, n = 816). Sequencing of rpoD gene and phylogenetic analysis revealed two aeromonads species, Aeromonas hydrophila and Aeromonas veronii. To the best of our knowledge this is the first report to establish the prevalence of aeromonads in apparently healthy farmed tilapia in Southern highlands and Northern zone of Tanzania. In addition it was observed that farmers were lacking proper knowledge and awareness on pond management practices and fish health management. In conclusion, the infection rate of aeromonads in apparently health tilapia coupled with lack of proper knowledge and awareness on pond and fish health management by fish farmers in the study area poses risk of diseases outbreaks in their farms in future. Therefore, it is recommended that the farmers should be trained on basic pond and fish health management and control strategies.

Ecological and Epidemiological Findings Associated with Zoonotic Rabies Outbreaks and Control in Moshi, Tanzania, 2017-2018.

Approximately 1500 people die annually due to rabies in the United Republic of Tanzania. Moshi, in the Kilimanjaro Region, reported sporadic cases of human rabies between 2017 and 2018. In response and following a One Health approach, we implemented surveillance, monitoring, as well as a mass vaccinations of domestic pets concurrently in >150 villages, achieving a 74.5% vaccination coverage (n = 29, 885 dogs and cats) by September 2018. As of April 2019, no single human or animal case has been recorded. We have observed a disparity between awareness and knowledge levels of community members on rabies epidemiology. Self-adherence to protective rabies vaccination in animals was poor due to the challenges of costs and distances to vaccination centers, among others. Incidence of dog bites was high and only a fraction (65%) of dog bite victims (humans) received post-exposure prophylaxis. A high proportion of unvaccinated dogs and cats and the relative intense interactions with wild dog species at interfaces were the risk factors for seropositivity to rabies virus infection in dogs. A percentage of the previously vaccinated dogs remained unimmunized and some unvaccinated dogs were seropositive. Evidence of community engagement and multi-coordinated implementation of One Health in Moshi serves as an example of best practice in tackling zoonotic diseases using multi-level government efforts. The district-level establishment of the One Health rapid response team (OHRRT), implementation of a carefully structured routine vaccination campaign, improved health education, and the implementation of barriers between domestic animals and wildlife at the interfaces are necessary to reduce the burden of rabies in Moshi and communities with similar profiles.

Current State of Modern Biotechnological-Based Aeromonas hydrophila Vaccines for Aquaculture: A Systematic Review.

This systematic review describes what "the cutting edge vaccines for Aeromonas hydrophila are". The focus is on types of high tech biotechnological based vaccines, target gene or antigen in developing these vaccines, and challenge model fish species used in vaccines efficacy testing. Vaccines delivery methods, immune response, and their efficacy, adjuvant or carrier systems used, and the overall experimental setup or design of the vaccines under investigation are also described. The search for the original papers published between 2009 and 2018 was conducted in June of 2018, using the PubMed and Google scholar electronic database. Twenty-three (23/4386) studies were included in the final assembly using PRISMA guidelines (Protocol not registered). Recombinant protein vaccines were the highly experimented type of the modern biotechnological based vaccines identified in the selected studies (16/23; 70%). Outer membrane proteins (OMPs) of different ß-barrels were shown to be a potential antigenic entity for A. hydrophila vaccines (57%). Intraperitoneal route with conventional carries or adjuvants was the highly applied delivery system while very few studies used herbal based vaccine adjuvants and nanomaterial as a vaccine carrier. Variation was observed in terms of protection levels in the selected studies. The experimental designs partly contributed to the observed variation. Therefore, recombinant vaccines that use new carrier system technologies and delivered through oral route in feeds would have been of great value for use in the prevention and control of A. hydrophila infections in fish. Despite the usefulness as academic tools to identify what is important in pathogenicity of the etiological agent to the host fish, these vaccines are only economically viable in very high-value animals. Therefore, if vaccination is a good option for A. hydrophila group, then simple autogenous vaccines based on accurate typing and evidence-based definition of the epidemiological unit for their use would be the most viable approach in terms of both efficacy and economic feasibility especially in low and middle-income countries (LMIC).

Taenia solium taeniosis and cysticercosis literature in Tanzania provides research evidence justification for control: A systematic scoping review.

BACKGROUND: Despite Taenia solium taeniosis/cysticercosis (TSTC) having been put high on the global agenda of neglected tropical diseases (NTDs), which over the last years has received a lot of attention, there has been no control programmes in place in sub-Saharan Africa, a highly endemic region. This could be attributed to lack of awareness of many stakeholders on the burden and impact of T. solium. This information is essential in guiding TSTC policies, practices and research agendas as well as encouraging cross-sectoral collaboration in the control of this important zoonotic parasite using a One Health approach. National elimination of the parasite is the foundation for global eradication. This will require that substantial country-level information is provided to all key stakeholders. We have mapped out TSTC research evidence in Tanzania to inform on disease burden and potential for integrated control measures. METHODOLOGY/PRINCIPAL FINDINGS: A scoping review of all TSTC studies undertaken in Tanzania and published up to December 2018 was conducted. The articles were searched from PUBMED, AJOL, Google Scholar and Google in general. Fifty-one (51) articles met the inclusion criteria and were reviewed. Prevalence of taeniosis of 2.3% - 5.2% was estimated based on copro-antigen ELISA while human cysticercosis of >16% was estimated based on serum antigen ELISA (Ag-ELISA) or IgG Western Blot. Neurocysticercosis (NCC) contributed significantly to epilepsy in adults. Farm prevalence of porcine cysticercosis were 6.0% - 17.4% (lingual examination) and 1.5% - 33.3% (Ag-ELISA). Slaughter-slab prevalence were 0% - 18.2% (routine meat inspection). Lacking latrines, watering pigs with river or pond water, and feeding pigs with potato peels were associated with porcine cysticercosis prevalence. Washing hands by dipping method increased the risk of human cysticercosis. In 2012, the number of DALYs/1000 person-years for NCC-associated epilepsy was 0.7 (95% UI, 0.2-1.6), around 5 million USD (95% UI, 797,535-16,933,477) were spent due to NCC-associated epilepsy and nearly 3 million USD (95% UI, 1,095,960-5,366,038) were potentially lost due to porcine cysticercosis. Three rounds of annual treatment of school-age children with praziquantel significantly reduced prevalence of taeniosis and porcine cysticercosis. Health education was efficacious in improving knowledge and attitudes favourable for control of TSTC while a single dose of oxfendazole 30 mg/kg body weight was efficacious in eliminating T. solium cysticerci from pig musculature. CONCLUSIONS/SIGNIFICANCE: The observed high burden of TSTC and the significant contribution of NCC to epilepsy in Tanzania warrant urgent interventions. Evaluation of best control options should make use of disease transmission dynamics models such as cystiSim, taking into account findings from the field based intervention studies. In addition, locally adapted management guidelines for people suffering from NCC are urgently needed.

Tilapia (Oreochromis niloticus) as a Putative Reservoir Host for Survival and Transmission of Vibrio cholerae O1 Biotype El Tor in the Aquatic Environment.

Studies have reported the occurrence of Vibrio cholerae in fish but little is known about the interaction between fish and toxigenic V. cholerae as opposed to phytoplankton, which are well-established aquatic reservoirs for V. cholerae. The present study determined the role of tilapia (Oreochromis niloticus) as a reservoir host for survival and transmission of V. cholerae in aquatic environments. Three experiments were performed with one repetition each, where O. niloticus (∼2 g) kept in beakers were inoculated with four V. cholerae strains (5 × 107 cfu/mL). Firstly, infected tilapia were kept in stagnant water and fed live brine shrimp (Artemia salina) larvae daily. Secondly, infected tilapia were kept without feeding and water was changed every 24 h. Thirdly, infected tilapia were fed and water was renewed daily. Infected tilapia and non-infected controls were sacrificed on days 1, 2, 3, 7, and 14 post-inoculation and V. cholerae were enumerated in intestinal content and water. Another experiment assessed the transmission of V. cholerae from infected to non-infected tilapia. The study revealed that El Tor biotype V. cholerae O1 and V. cholerae non-O1 colonized tilapia intestines and persisted at stable concentrations during the second week of the experiment whereas the Classical biotype was undetectable after 1 week. In stagnant water with feeding, V. cholerae counts dropped to 105 cfu/ml in water and from 107 to 104 cfu/intestine in fish after 14 days. When water was renewed, counts in water decreased from 107 to 103 cfu/ml and intestinal counts went from 106 to 102 cfu/intestine regardless of feeding. All strains were transmitted from infected to naïve fish after 24 h of cohabitation. Tilapia like other fish may play an essential role in the survival and dissemination of V. cholerae O1 in aquatic environments, e.g., the seventh pandemic strains mostly. In this study, tilapia were exposed to high concentrations of V. cholerae to ensure initial uptake and follow-up studies with lower doses resembling natural concentrations of V. cholerae in the aquatic environment are needed to confirm our findings.

Surveillance and Genomics of Toxigenic Vibrio cholerae O1 From Fish, Phytoplankton and Water in Lake Victoria, Tanzania.

The occurrence of toxigenic Vibrio cholerae O1 during a non- outbreak period in Lake Victoria was studied and genetic characteristics for environmental persistence and relatedness to pandemic strains were assessed. We analyzed 360 samples of carps, phytoplankton and water collected in 2017 during dry and rainy seasons in the Tanzanian basin of Lake Victoria. Samples were tested using PCR (ompW and ctxA) with DNA extracted from bacterial isolates and samples enriched in alkaline peptone water. Isolates were screened with polyvalent antiserum O1 followed by antimicrobial susceptibility testing. Whole genome sequencing and bioinformatics tools were employed to investigate the genomic characteristics of the isolates. More V. cholerae positive samples were recovered by PCR when DNA was obtained from enriched samples than from isolates (69.0% vs. 21.3%, p < 0.05), irrespectively of season. We identified ten V. cholerae O1 among 22 ctxA-positive isolates. Further studies are needed to serotype the remaining ctxA-positive non-O1 strains. Sequenced strains belonged to El Tor atypical biotype of V. cholerae O1 of MLST ST69 harboring the seventh pandemic gene. Major virulence genes, ctxA, ctxB, zot, ace, tcpA, hlyA, rtxA, ompU, toxR, T6SS, alsD, makA and pathogenicity islands VPI-1, VPI-2, VSP-1, and VSP-2 were found in all strains. The strains contained Vibrio polysaccharide biosynthesis enzymes, the mshA gene and two-component response regulator proteins involved in stress response and autoinducers for quorum sensing and biofilm formation. They carried the SXT integrative conjugative element with phenotypic and genotypic resistance to aminoglycoside, sulfamethoxazole, trimethoprim, phenicol, and quinolones. Strains contained a multidrug efflux pump component and were resistant to toxic compounds with copper homeostasis and cobalt-zinc-cadmium resistance proteins. The environmental strains belonged to the third wave of the seventh pandemic and most are genetically closely related to recent outbreak strains from Tanzania, Kenya, and Uganda with as low as three SNPs difference. Some strains have persisted longer in the environment and were more related to older outbreak strains in the region. V. cholerae O1 of outbreak potential seem to persist in Lake Victoria through interactions with fish and phytoplankton supported by the optimum water parameters and intrinsic genetic features enhancing survival in the aquatic environment.

Effect of clove (Syzygium aromaticum) and seaweed (Kappaphycus alvarezii) water extracts pretreatment on lipid oxidation in sun-dried sardines (Rastrineobola argentea) from Lake Victoria, Tanzania.

Small indigenous fish species play a significant role in food and nutritional security of poor communities in developing countries. Sardines (Rastrineobola argentea) are fish species of Lake Victoria known to be a good source of health-promoting omega-3 fatty acids. Open sun drying is a common and traditional sardine processing and preservation method. Sun-dried products suffer from characteristic off-flavor due to lipid oxidation which discourage product consumption and limit diversification. This study investigated the use of clove (Syzygium aromaticum) and seaweed (Kappaphycus alvarezii) water extracts as natural antioxidants to impede lipid oxidation in sun-dried sardines. Lipid oxidation was assessed by peroxide value, volatile secondary oxidation products, and fatty acid profiles. The antioxidant capacity of extracts was evaluated by total phenolic content, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, and iron (Fe2+) chelating ability. Results showed that 5, 10, and 20 g/L clove extracts significantly reduced peroxidation in sun-dried sardines by 38.7%, 54.6%, and 56%, respectively. Clove extracts resulted in higher retention of omega-3 fatty acids and lower concentrations of secondary lipid oxidation products as opposed to seaweed counterpart. This research has demonstrated feasibility of pretreating whole, omega-3-rich small sardines with natural antioxidants to avert lipid oxidation during sun drying.

Heavy metals in farmed and wild milkfish (Chanos chanos) and wild mullet (Mugil cephalus) along the coasts of Tanzania and associated health risk for humans and fish.

In 2016, farmed milkfish (Chanos chanos) from Tanzania mainland (Mtwara), and Zanzibar islands (Pemba and Unguja) and wild milkfish and mullet (Mugil cephalus) from the Indian Ocean were collected for analyses of heavy metals (Pb, Cd, Hg, As, Al, Fe, Zn, Cu, Ni, Co and Cr) in muscles and livers. High concentrations of Pb were detected in muscles and livers from wild and farmed milkfish and wild mullet from all sites. The highest concentration of Pb was detected in wild milkfish liver from Mtwara (47.4 mg/kg ww). The Pb concentrations in fish muscle exceeded maximum levels (ML) set by FAO/WHO (0.3 mg/kg ww) in 100% of the analysed fish. Concentrations of Pb were higher in wild fish than in farmed fish. Cd concentrations were generally low. The comparison of the Hg concentration with EQSBiota indicated that Hg might pose potential health risk to 22% of the analysed fish. Median concentrations of Fe in livers from farmed milkfish from Jozani and Shakani, Zanzibar, were 40-80 times higher than the other sites. Assessment of human health risk and exposure to heavy metals indicated no potential risk from consuming the fish from the present study locations. However, the Pb concentrations exceeding ML in the fish suggests that Pb may affect the health of fish. Future investigations should include regular monitoring of heavy metals in farmed and wild fish in Tanzania for further development of sustainable aquaculture and the welfare of the wild fish stock in the coastal waters.