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The early diagnosis of mastitis is an essential factor for the prompt detection of the animal for further actions. In fact, if not culled, infected cows must be segregated from the milking herd and milked last, or milked with separate milking units. Besides microbiological analysis, the somatic cell count (SCC) commonly used as predictor of intramammary infection, frequently lead to a misclassification of milk samples. To overcome these limitations, more specific biomarkers are continuously evaluated. The total amino acid content increases significantly in mastitic milk compared to normal milk. S. aureus requires branched-chain amino acids (BCAAs—isoleucine, leucine, and valine) for protein synthesis, branched-chain fatty acids synthesis, and environmental adaptation by responding to their availability via transcriptional regulators. The increase of BCAAs in composite milk has been postulated to be linked to mammary infection by S. aureus. The aim of this work is to demonstrate, by a direct ion-pairing reversed-phase method, based on the use of the evaporative light-scattering detector (IP-RP-HPLC-ELSD), applied to 65 composite cow milk samples, a correlation between the concentration of isoleucine and leucine, and S. aureus load. The correlation coefficient, r, was found to be 0.102 for SCC (p = 0.096), 0.622 for isoleucine (p < 0.0001), 0.586 for leucine (p < 0.0001), 0.013 for valine (p = 0.381), and 0.07 for tyrosine (p = 0.034), standing for a positive correlation between S. aureus and isoleucine and leucine concentration. The link between the content of BCAAs, isoleucine and leucine, and udder infection by S. aureus demonstrated with our study has an important clinical value for the rapid diagnosis of S. aureus mastitis in cows.

The prevalence of HIV/AIDS among men who have sex with men (MSM) in Nairobi has been reported to be 16.4% which is a public health concern. The aim of this study was to determine factors that affected accessibility to, acceptability of, and adherence to HIV/AIDS prevention services by men who have sex with men in Nairobi County, Kenya. Methods. The study used a mixed methods design, applying both quantitative and qualitative methods. Results. Over 70% of MSM reported that TB drugs, antiretroviral drugs, lubricants, condoms, and other STI treatment services were accessible. However, 31.6% of MSM reported that discrimination at the health facility level was a hindrance to accessing HIV/AIDS prevention services while 26.3% reported encountering stigma. 48.4% of MSM consumed alcohol while taking of drugs was reported, at 36.1%. Conclusion. Discrimination and stigma adversely affected MSM seeking HIV/AIDS prevention services in health facilities. MSM engaged in alcohol and substance abuse that likely affected their health seeking behavior. Although most prevention services are reported as accessible, uptake of the same needs to be considered.

The aim of this study was to monitor and evaluate the seasonal changes on selected reproductive parameters in mares. Ninety-six mares of different breeds, aged 3 to 23 years were evaluated during the breeding season 2015–2018 at the Equine clinic at University of Veterinary Medicine and Pharmacology in Košice, Slovakia (48°42´N, 21°15´E). The beginning of the estrus was determined by history or observation, mares were examined every 6 hours, blood for progesterone analysis was taken from the jugular vein. Correlation analyses were performed using both the Pearson and the Spearman correlation coefficient. Statistical analysis was performed using the functions of Microsoft Excel and GraphPad Prism. The day length (r = 0.708, P

The Dark Energy Spectroscopic Instrument (DESI) embarked on an ambitious 5 yr survey in 2021 May to explore the nature of dark energy with spectroscopic measurements of 40 million galaxies and quasars. DESI will determine precise redshifts and employ the baryon acoustic oscillation method to measure distances from the nearby universe to beyond redshift z > 3.5, and employ redshift space distortions to measure the growth of structure and probe potential modifications to general relativity. We describe the significant instrumentation we developed to conduct the DESI survey. This includes: a wide-field, 3.°2 diameter prime-focus corrector; a focal plane system with 5020 fiber positioners on the 0.812 m diameter, aspheric focal surface; 10 continuous, high-efficiency fiber cable bundles that connect the focal plane to the spectrographs; and 10 identical spectrographs. Each spectrograph employs a pair of dichroics to split the light into three channels that together record the light from 360–980 nm with a spectral resolution that ranges from 2000–5000. We describe the science requirements, their connection to the technical requirements, the management of the project, and interfaces between subsystems. DESI was installed at the 4 m Mayall Telescope at Kitt Peak National Observatory and has achieved all of its performance goals. Some performance highlights include an rms positioner accuracy of better than 0.″1 and a median signal-to-noise ratio of 7 of the [O ii ] doublet at 8 × 10 ^−17 erg s ^−1 cm ^−2 in 1000 s for galaxies at z = 1.4–1.6. We conclude with additional highlights from the on-sky validation and commissioning, key successes, and lessons learned.

Since ancient times the concept of dose response, from a toxicological perspective, has been a matter of concern. Already by the 8th century BC and over the years, many enlightened people have attempted to interpret this phenomenon, observing and coming across its results and practical implementation through exposure to chemical substances, either from natural or synthetic sources. Nowadays, the environmental exposure of human populations to chemicals in terms of quantity and quality might differ. Nevertheless, dose response still remains an issue joining hands with scientific and technological progress. The aim of the present review is not only to briefly recount the history of the dose response concept, from ancient time theories to novel approaches, but also to draw the outline of challenges and requirements toxicology science needs to fulfill. Keywords: Dose response, Mithridatism, Tolerance, Hormesis, Risk assessment, Biomonitoring, Toxicology science

Fertility problems of mares on a well-managed breeding farm with thoroughbred stallions have been ascribed mostly to contamination of the reproductive apparatus of females with pathogens, particularly those of bacterial origin. This study presents a summary of the frequency of bacterial pathogens isolated from 437 cervical swabs which were collected from English thoroughbred mares intended for mating between 2008-2014, as well as of resistance tests of these pathogens to seven commonly used antibiotics as follows: penicillin, gentamicin, tetracycline, sulfisoxazole, cefotaxime, marbofloxacin and enrofloxacin. In addition, the study reports the changes in the levels of plasma oestradiol and progesterone determined two to three days before and two to three days after the first post-partum ovulation in mares with positive and dubious bacteriological findings and percentage of barren mares and mares that conceived at first, second and third post-partum ovulations. It was observed that 21.5% of mares were barren even after the third post-partum cycles. The oestradiol levels determined two to three days before the first post-partum ovulation were significantly lower in mares positive for pathogenic microflora in their reproductive apparatus compared to mares with the dubious findings (25.1 ± 5.8 pg/ml vs. 69.7 ± 18.3 pg/ml; P < 0.05), while the mean progesterone levels did not differ significantly but displayed a rather wide range in positive mares (from 0.08 to 1.38 ng/ml) compared to dubious mares with only small variations (0.12 ± 0.03 ng/ml). Moreover, of the total number of cervical swabs taken shortly before the first post-partum oestrus from all the mares intended for mating as many as 69.7% were contaminated with pathogenic microflora (positive findings). Saprophytic microorganisms only (the dubious findings) were isolated from 29.7% of swabs. From the 307 positive swabs, we could identify 40.4% positive for β-haemolytic streptococci and 20.4% positive for Escherichia coli, the pathogens implicated in causing reproductive disorders. Tests of antibiotic resistance of the investigated pathogens revealed that both Gram-positive and Gram-negative bacteria showed high susceptibility to antibiotics such as cefotaxime, marbofloxacin and enrofloxacin. On the other hand, both these bacterial groups showed high resistance to routinely used broad-spectrum antibiotics, such as penicillin and tetracycline. Because further research is required for a full understanding of the mechanism of pathogenesis of post-breeding endometritis, we can only hypothesise that uterine contamination with pathogenic microflora, particularly with β-haemolytic streptococci and coliform bacteria, diagnosed before the first post-partum ovulation, could negatively affect the hormonal regulation of oestrus and result in mare fertility problems.

The Dutch merchant and naturalist Anton van Leeuwenhoek is considered to be the father of optic microscopy and the precursor of bacteriology. Among others, he discovered and studied the spermatozoon.El comerciante y biólogo holandés Anton van Leeuwenhoek es considerado el padre de la microscopía óptica y el precursor de la bacteriología. Entre otros, descubrió y entendió a los espermatozoides.

In this report, a case of malignant fibrous histiocytoma involving skin, lungs, kidneys, pancreas and mediastinal lymph node was described. Microscopically, the tumor classified as storiform-pleomorphic type malignant fibrous histiocytoma.

Phenolic compounds were extracted with 80% methanol from caryopses and embryos of rye (cv. Dańkowskie Złote and Amilo). In all extracts, reducing power, scavenging effect on DPPH radical, and antioxidant activity in a β-carotene-linoleate model system were examined. The highest content of total phenolic compounds was noted in the extract from caryopses of Amilo (7.93 mg/g of extract). UV spectra of all extracts were characterised by maxima originated from phenolic acids (320, 326 and 328 nm), and by maxima at shorter wavelengths (272 and 274 nm) attributed to other phenolic compounds. All extracts showed a good antioxidant activity in a β-carotene-linoleate model system. This activity was similar to that reported before in leguminous seeds extracts. The antioxidant activities of the extracts from the caryopses of Dańkowskie Złote and the embryos of both cultivars were very similar, especially during the second part of the incubation period. The extract of Amilo embryos showed a slightly weaker antioxidative effect. The weak antiradical effects in the experiments with DPPH radical and a weak reducing power were characteristic for all the extracts investigated.

Two pea protein isolates, Pisane and Propulse, were hydrolysed by trypsin. The degree of hydrolysis (DH) was computed using a pH-stat method. Enzymatic treatment of the pea protein isolates was conducted at four different temperatures, namely 35, 40, 45 and 50°C. The relationship between DH and E/S ratio was studied at 50°C and at four different E/S ratios; these were 5, 15, 25, 35 mAU/g (AU - Anson unit). For Pisane the highest value of the final DH (10.4%) was obtained at 45°C, whereas for Propulse the optimal temperature was 50°C and a DH value of 13.2% was attained. In the case of Pisane, the highest DH (11.5%) was recorded if the enzyme/substrate ratio was 35 mAU/g whereas for Propulse, the highest DH (13.2%) was observed at an E/S ratio of 15 mAU / g.

An extract of seeds of pea was prepared using 80% (v/v) acetone. Six fractions (I-VI) were separated from the crude extract on a Sephadex LH-20 column using methanol as the mobile phase. The antioxidant activity of fractions was investigated in a β-carotene-linoleate model system. For individual fractions, UV spectra were recorded and the content of total phenolics was determined. Results of the β-carotene-linoleate model system indicated that antioxidant activities of separated fractions correlated with their content of total phenolic compounds and they decreased in the order of IV = VI > V > III > I > II. The antioxidant activity of fractions IV and VI was very strong as compared with that of butylated hydroxyanisole (BHA). Absorption maxima from UV spectra showed that flavonoids, and not phenolic acids, were the main phenolic compounds in separated fractions.

Phenolic compounds were extracted from lentil seeds using three solvent systems: 80% (v/v) acetone, 80% (v/v) methanol, and 80% (v/v) ethanol. Each extract was subsequently separated into two fractions by chromatoghraphy on a column with Toyo Pearl HW-40 using water (fraction I) and methanol (fraction II) for elution. Antioxidative activity of extracts and their respective fractions were examined in a β-carotene-linoleate model system. All three extracts exhibited similar antioxidant activity. Considering the level of phenolic compounds in extracts it seems that phenolic compounds from the acetone extract were less active than those from either the methanolic and ethanolic ones. Because the content of phenolics was about 16-fold lower in fraction I of the methanolic and ethanolic extracts compared to fraction II, the phenolics in fraction I of the methanol and ethanol extracts from lentil seeds are much more active than these in fraction II. A stronger antioxidant activity of fraction I from the acetone extract compared to the crude acetone extract was observed during the latter incubation stage. The reason was a relatively high level of phenolic compounds in this fraction. UV spectra confirmed that the phenolic compounds from the acetone extract were different compared to methanolic and ethanolic extracts.

In model experiments with the use of Miller's test to 5 ml emulsion 25, 12.5, 6.25 and 3.13µg BHA, BHT and TBHQ was added. Inhibitory ratios (IR) for BHA and BHT were similar. At addition level of 6.25 and 3.13 IR for TBHQ was lower than for BHA and BHT. It is suggested that in the model studies with the use of Miller's test 25 µg BHA, BHT or TBHQ should be added to 5 ml of control sample.

Simon-Emmanuel Duplay was a distinguished example of the French clinicopathological school and his researches on cancer inaugurated the modern period in oncology. In his book entitled Les tumeurs, he presents and analyses his research and those of his contemporaries on cancer contagion.

Four thousand years ago the willow tree was known for its palliative effects. Ancient Greeks used the extract from the cortex and leaves to treat pain, fever, calluses and to enhance semen quality. Bayer Company and Felix Hoffmann claimed discovery of aspirin, and chemists all over Europe dealt with the synthesis of the drug. During 1988 the use of Aspirin was extended from an analgesic, antipyretic to a life-saving drug as it was suggested that it reduced the danger of cardiovascular accidents. Aspirin is still a drug that can offer much to medicine.

Protein-polyphenol association may affect polyphenols absorption/bioavailability from gastrointestinal tract and finally modulate their biological activities. Fluorescence spectroscopy has been widely used to investigate the interactions of drugs and proteins. Measuring of quenching of intrinsic fluorescence of proteins can be also applied to study binding affinity of phenolic compounds to proteins. The aim of our study was to investigate interactions between phenolic compounds and proteins (isolated from broad bean seeds) using fluorescence quenching method. Phenolic compounds were extracted from broad bean coats and separated chromatographically into low molecular weight (LMW) fraction and tannins. Total phenolic contents of low molecular weight and tannin fractions were 203.6 and 740.5 mg of catechin equivalents per mg, respectively. Protocatechuic, gallic, p-hydroxybenzoic, p-coumaric acids and epicatechin were identified as predominant constituents of low molecular weight fraction using RP-HPLC method. Column chromatography on Sephadex G-200 gel was used for purification of 11S globulins from broad bean seeds. Fluorometric titration was applied to measure quenching of 11S globulin fluorescence by phenolic compounds. Titration curves depict dose dependent decrease of fluorescence intensity of protein both for LMW fraction and tannins. However, tannin fraction was more effective quencher of protein fluorescence than low molecular weight fraction. Emission spectra of 11S globulin at λex 282 nm showing quenching effect of phenolic compounds were recorded. Additionally, binding constants were calculated to characterise binding affinity of p-hydroxybenzoic acid and epicatechin to 11S protein using Stern-Volmer plot. The results were corrected both for inner filter effect and phenolic compounds intrinsic fluorescence.

Rapeseeds of 6.5, 8.2, 9.9 and 12.65% moisture were pressed to obtain oil. After two weeks methanol extract of phenolic compounds was obtained from the oil cake. Analysis reveal that there was not effect of seed moisture on the content of total phenolic compounds in extracts and on antioxidative activity of extracts. There were no significant differences in UV spectra or diagrams of derivatives 2 and 4 between the extracts.

Antimicrobial resistance (AMR) poses a significant global One Health challenge that causes increased mortality and a high financial burden. Animal production contributes to AMR, as more than half of antimicrobials are used in food-producing animals globally. There is a growing body of literature on AMR in food-producing animals in African countries, but the surveillance practices across countries vary considerably. This pilot study aims to explore the potential of wastewater and environmental surveillance (WES) of AMR and its extension to the veterinary field. Floor drainage swab ( n  = 18, 3/abattoir) and wastewater ( n  = 16, 2-3/abattoir) samples were collected from six South African abattoirs that handle various animal species, including cattle, sheep, pig, and poultry. The samples were tested for Extended-Spectrum Beta-Lactamase (ESBL) and Carbapenemase-producing Enterobacterales, Methicillin-Resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Enterococci (VRE), and Candida auris by using selective culturing and MALDI-TOF MS identification. The phenotype of all presumptive ESBL-producing Escherichia coli ( n  = 60) and Klebsiella pneumoniae ( n  = 24) isolates was confirmed with a disk diffusion test, and a subset (15 and 6 isolates, respectively), were further characterized by whole-genome sequencing. In total, 314 isolates (0-12 isolates/sample) withstood MALDI-TOF MS, from which 37 species were identified, E. coli and K. pneumoniae among the most abundant. Most E. coli ( n  = 48/60; 80%) and all K. pneumoniae isolates were recovered from the floor drainage samples, while 21 presumptive carbapenem-resistant Acinetobacter spp. isolates were isolated equally from floor drainage and wastewater samples. MRSA, VRE, or C. auris were not found. All characterized E. coli and K. pneumoniae isolates represented ESBL-phenotype. Genomic analyses revealed multiple sequence types (ST) of E. coli ( n  = 10) and K. pneumoniae ( n  = 5), including STs associated with food-producing animals globally, such as E. coli ST48 and ST10 and K. pneumoniae ST101. Common beta-lactamases linked to food-producing animals, such as bla CTX-M-55 and bla CTX-M-15 , were detected. The presence of food-production-animal-associated ESBL-gene-carrying E. coli and K. pneumoniae in an abattoir environment and wastewater indicates the potential of WES in the surveillance of AMR in food-producing animals. Furthermore, the results of this pilot study encourage studying the topic further with refined methodologies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Heljanko, Karama, Kymäläinen, Kurittu, Johansson, Tiwari, Nyirenda, Malahlela and Heikinheimo.)

Nontyphoidal Salmonella enterica serovars are foodborne pathogens commonly transmitted through poultry products. Draft genome sequences of three Salmonella enterica subsp. enterica serovar Shamba isolates which were obtained from poultry house dust in South Africa are reported herein., Competing Interests: The authors declare no conflict of interest.

Shiga toxin-producing and Enteropathogenic Escherichia coli are foodborne pathogens commonly associated with diarrheal disease in humans. This study investigated the presence of STEC and EPEC in 771 dairy cattle fecal samples which were collected from 5 abattoirs and 9 dairy farms in South Africa. STEC and EPEC were detected, isolated and identified using culture and PCR. Furthermore, 339 STEC and 136 EPEC isolates were characterized by serotype and major virulence genes including stx1, stx2, eaeA and hlyA and the presence of eaeA and bfpA in EPEC. PCR screening of bacterial sweeps which were grown from fecal samples revealed that 42.2% and 23.3% were STEC and EPEC positive, respectively. PCR serotyping of 339 STEC and 136 EPEC isolates revealed 53 different STEC and 19 EPEC serotypes, respectively. The three most frequent STEC serotypes were O82:H8, OgX18:H2, and O157:H7. Only 10% of the isolates were classified as "Top 7" STEC serotypes: O26:H2, 0.3%; O26:H11, 3.2%; O103:H8, 0.6%; and O157:H7, 5.9%. The three most frequent EPEC serotypes were O10:H2, OgN9:H28, and O26:H11. The distribution of major virulence genes among the 339 STEC isolates was as follows: stx1, 72.9%; stx2, 85.7%; eaeA, 13.6% and hlyA, 69.9%. All the 136 EPEC isolates were eaeA-positive but bfpA-negative, while 46.5% carried hlyA. This study revealed that dairy cattle are a major reservoir of STEC and EPEC in South Africa. Further comparative studies of cattle and human STEC and EPEC isolates will be needed to determine the role played by dairy cattle STEC and EPEC in the occurrence of foodborne disease in humans.Please kindly check and confirm the country and city name in affiliation [6].This affiliation is correct.Please kindly check and confirm the affiliationsConfirmed. All Affiliations are accurate., (© 2024. The Author(s).)

Salmonella enterica is a zoonotic pathogen and a leading cause of foodborne gastroenteritis in humans. Here, we report the draft genome sequences of two Salmonella Uzaramo isolates, which were isolated from poultry organs during routine post-mortem examination in South Africa. Currently, whole-genome sequences on Salmonella Uzaramo are scanty., Competing Interests: The authors declare no conflict of interest.

In this study, four antimicrobial growth promoters, including virginiamycin, josamycin, flavophospholipol, poly 2-propenal 2-propenoic acid and ultraviolet light, were tested for their capacity to induce stx-bacteriophages in 47 Shiga toxin-producing E. coli O157:H7 isolates. Induced bacteriophages were characterized for shiga toxin subtypes and structural genes by PCR, DNA restriction fragment length polymorphisms (RFLP) and morphological features by electron microscopy. Bacteriophages were induced from 72.3% (34/47) of the STEC O157:H7 isolates tested. Bacteriophage induction rates per induction method were as follows: ultraviolet light, 53.2% (25/47); poly 2-propenal 2-propenoic acid, 42.6% (20/47); virginiamycin, 34.0% (16/47); josamycin, 34.0% (16/47); and flavophospholipol, 29.8% (14/47). A total of 98 bacteriophages were isolated, but only 59 were digestible by NdeI, revealing 40 RFLP profiles which could be subdivided in 12 phylogenetic subgroups. Among the 98 bacteriophages, stx2a, stx2c and stx2d were present in 85.7%, 94.9% and 36.7% of bacteriophages, respectively. The Q, P, CIII, N1, N2 and IS1203 genes were found in 96.9%, 82.7%, 69.4%, 40.8%, 60.2% and 73.5% of the samples, respectively. Electron microscopy revealed four main representative morphologies which included three bacteriophages which all had long tails but different head morphologies: long hexagonal head, oval/oblong head and oval/circular head, and one bacteriophage with an icosahedral/hexagonal head with a short thick contractile tail. This study demonstrated that virginiamycin, josamycin, flavophospholipol and poly 2-propenal 2-propenoic acid induce genetically and morphologically diverse free stx-converting bacteriophages from STEC O157:H7. The possibility that these antimicrobial growth promoters may induce bacteriophages in vivo in animals and human hosts is a public health concern. Policies aimed at minimizing or banning the use of antimicrobial growth promoters should be promoted and implemented in countries where these compounds are still in use in animal agriculture., (© 2023. The Author(s).)

Community-led total sanitation (CLTS) is a widely used approach for enhancing sanitation practices. However, the impact of boosted CLTS on household latrine ownership has not been adequately evaluated. This study aims to investigate the factors associated with latrine possession among households, with a specific focus on single and CLTS-boosting implementation. A community-based repeated cross-sectional study was conducted in Siaya County, Kenya, involving 512 households at the baseline and 423 households at the follow-up. Data were analyzed using the mixed-effects logistic regression model. At the baseline, latrine possession was significantly associated with CLTS implementation (adjusted OR [aOR]: 3.01; 95% confidence interval [CI]: 1.41-6.44), literacy among households (aOR: 1.83; 95% CI: 1.12-2.98) and higher socioeconomic status (SES) (second level: aOR: 2.48; 95% CI:1.41-4.36, third level: aOR: 3.11; 95% CI: 1.76-5.50, fourth level: aOR: 10.20; 95% CI: 5.07-20.54). At follow-up, CLTS boosting (aOR: 7.92; 95% CI: 1.77-35.45) and a higher SES were associated with increased latrine ownership (second level: aOR: 2.04; 95% CI: 0.97-4.26, third level: aOR: 7.73; 95% CI: 2.98-20.03, fourth level: aOR: 9.93; 95% CI: 3.14-28.35). These findings highlight the significant role played by both single and CLST boosting in promoting universal latrine ownership and empowering vulnerable households to understand the importance of sanitation and open defecation-free practices.

The latest EU regulation on geographical indications (EU Regulation No. 1151/2012) has introduced a set of new tools for the protection and enhancement of food products in rural areas, under the group name of optional quality term (OQT). The Commission Delegated EU Regulation, No. 665/2014, regulated the conditions for the use of the optional quality term mountain product (MP), to support the implementation of a mountain value chain. This new tool is aimed at promoting local development, maintaining the economic activities in mountain areas, and redistributing wealth, whilst, at the same time, promoting the territory. Pecorino and goat cheeses are typical Italian cheeses made usually with whole raw ewe's or raw goat's milk, without starter culture addition. In an attempt to characterize these productions, the aim of this study was to investigate the evolution of enterococci during the production and ripening of Pecorino cheese made in three different farms, located in Umbria, Italy in areas facing natural or other specific constraints as stipulated by Regulation 1305/2013 on support for rural development by the European Agricultural Fund for Rural Development (EAFRD). Enterococci are enteric organisms which are commonly isolated from ewe and goat's milk production in Umbria, Italy. Counts of enterococci in raw milk ranged from 1.75 for ovine milk to 3.62 for ewe milk and a marked reduction was observed after thermization especially in ovine milk. Out of 100 isolates, 69 were E. faecium , 23 E. durans , 8 E. faecalis and 2 E. casseliflavus and the distribution of species between farms and between samples showed a prevalence of E. faecium in ovine farms and E. durans in ewes farms, with an equal dis-tribution between samples. High percentages of susceptible isolates were found for amoxicil-lin/clavulanic acid, ampicillin, chloramphenicol, sulphamethoxazole, sulphamethoxazole/ trimethoprim, ticarcillin, vancomycin. A high prevalence of resistant strains (>30%) was ob-served for amikacin, ciprofloxacin, ceftriaxone, kanamycin, tetracycline. A comparison of this re-sults with those of previous works on similar dairy products revealed high levels of resistance to antimicrobials which needs to be addressed., Competing Interests: Conflict of interest: The authors declare no potential conflict of interest., (©Copyright: the Author(s).)

Packaging is considered one of the most interesting technological aspects of food production and is a constantly evolving subject in food production. The type of packaging is important for the quality and safety of the product and for the visual appearance of the product to be immediately evaluated by consumers. The purpose of this study was to investigate the effect of four different types of modified atmosphere packaging (ATM) and vacuum packaging (VP) currently used by a company in central Italy, on the main qualitative characteristics of beef. For these two traditional and two new solutions with reduced environmental impact and compostable were evaluated. For each type of packaging, two different products were analyzed: steaks and hamburgers. The samples, immediately after production, were transported to the laboratory in refrigerated containers. Several parameters (color, pH, water holding capacity, drip loss, and microbiological characteristics) were evaluated at time 0 and after 7 (T7), 14 (T14) and 21 days (T21) of storage in the dark and at refrigeration temperature (+4°C ± 2°C). The results showed that the two types of packaging have very similar effects on the water-retaining capacity of the steaks. More noticeable differences were recorded by the colorimetric analysis: for both steaks and hamburgers, the products packaged in the traditional packaging appeared brighter and redder than those packaged in the new alternatives. The microbiological analysis of the steaks showed higher values in the "new" packaging. The formation of abundant ropy slime was observed in one of the samples in the "new" modified atmosphere package at T21. The results of this study showed that the technological characteristics (in particular, the color) and the microbiological characteristics of the steaks and hamburgers were better in "old" packaging, with a better appearance and a longer shelf life. The results obtained show how the research for eco-sustainable products for packaging must be addressed, taking into account the effect of the materials on the qualitative and hygienic- sanitary characteristics of the meat., Competing Interests: Conflict of interest: The authors declare no potential conflict of interest., (©Copyright: the Author(s).)

Abstract: Staphylococcus pseudintermedius ( SP ) is an important opportunistic pathogen, frequently associated with pyoderma and otitis in dogs. The emergence and rapid expansion of methicillin-resistant Staphylococcus pseudintermedius (MR SP ) is problematic due to multidrug resistance and reduced treatment options. The aim of this study was to determine i) the prevalence of MR SP in dogs with pyoderma or otitis externa, ii) the antimicrobial resistance patterns of MR SP from South African isolates, and iii) the risk factors for MR SP -associated pyoderma or otitis externa in dogs in South Africa (RSA). Sixty-eight presumptive clinical SP isolates (collected from 65 dogs) from five geographically dispersed laboratories in RSA were collected over 2 years. Possible MR SP isolates were flagged when resistance to oxacillin was observed. Thereafter, all isolates were confirmed as SP by polymerase chain reaction (PCR) and further genotyped for the mec A gene. Fifty-seven of 68 isolates were confirmed to be SP (83.8%), while 49/57 (85.9%) carried mec A. Our findings showed that preliminary phenotypic methods supplemented by genotypic methods increased the accuracy of correctly identifying SP . All isolates were resistant to at least one antimicrobial drug. There was a high incidence of amoxicillin (70.1%) and enrofloxacin (65%) resistance. Important risk factors for mec A positive carriage were previous hospital admission, pruritus, and previous antibacterial failure. This study demonstrates a high prevalence of mec A positive carriage (85.9% of samples) in MR SP pyoderma and otitis in dogs in RSA. There is an urgent need for better laboratory diagnosis of MR SP and surveillance of dogs presenting with pyoderma and otitis in South Africa., (Copyright© Authors.)

Shiga-toxin-producing Escherichia coli is a foodborne pathogen commonly associated with human disease characterized by mild or bloody diarrhea hemorrhagic colitis and hemolytic uremic syndrome. This study investigated the occurrence of STEC in fecal samples of 289 goats in South Africa using microbiological culture and PCR. Furthermore, 628 goat STEC isolates were characterized by serotype (O:H) and major virulence factors by PCR. STEC was found in 80.2% (232/289) of goat fecal samples. Serotyping of 628 STEC isolates revealed 63 distinct serotypes including four of the major top seven STEC serogroups which were detected in 12.1% (35/289) of goats: O157:H7, 2.7% (8/289); O157:H8, 0.3%, (1/289); O157:H29, 0.3% (1/289); O103:H8, 7.6% (22/289); O103:H56, 0.3% (1/289); O26:H2, 0.3% (1/289); O111:H8, 0.3% (1/289) and 59 non-O157 STEC serotypes. Twenty-four of the sixty-three serotypes were previously associated with human disease. Virulence genes were distributed as follows: stx 1, 60.6% (381/628); stx 2, 72.7% (457/628); eaeA , 22.1% (139/628) and hlyA , 78.0% (490/628). Both stx 1 and stx 2 were found in 33.4% (210/628) of isolates. In conclusion, goats in South Africa are a reservoir and potential source of diverse STEC serotypes that are potentially virulent for humans. Further molecular characterization will be needed to fully assess the virulence potential of goat STEC isolates and their capacity to cause disease in humans.

Tungiasis is a cutaneous parasitosis caused by an embedded female sand flea. The distribution of cases can be spatially heterogeneous even in areas with similar risk profiles. This study assesses household and remotely sensed environmental factors that contribute to the geographic distribution of tungiasis cases in a rural area along the Southern Kenyan Coast. Data on household tungiasis case status, demographic and socioeconomic information, and geographic locations were recorded during regular survey activities of the Health and Demographic Surveillance System, mainly during 2011. Data were joined with other spatial data sources using latitude/longitude coordinates. Generalized additive models were used to predict and visualize spatial risks for tungiasis. The household-level prevalence of tungiasis was 3.4% (272/7925). There was a 1.1% (461/41,135) prevalence of infection among all participants. A significant spatial variability was observed in the unadjusted model ( p -value < 0.001). The number of children per household, earthen floor, organic roof, elevation, aluminum content in the soil, and distance to the nearest animal reserve attenuated the odds ratios and partially explained the spatial variation of tungiasis. Spatial heterogeneity in tungiasis risk remained even after a factor adjustment. This suggests that there are possible unmeasured factors associated with the complex ecology of sand fleas that may contribute to the disease's uneven distribution.

The immediate refrigeration of meat after slaughter is a key issue for the proper storage and aging of meat. The industry standard cold chain relies on low temperatures and ventilation to lower the internal carcass temperature to 0-4 °C within the first 48 h, i.e., within four times the so-called semi-cooling time. On the other hand, for games, once bled and eviscerated, the carcass must be sent to a point where it can be sectioned or kept on air for maturation at refrigeration temperature. The precautions to observe are few and simple but essential: protect the meat and start the cooling process quickly. After preparing the animal (bleeding and evisceration), it may be necessary to face a period of transport that is sometimes long and not very easy; while small animals can be easily transported in a backpack, larger ones must necessarily be carried by several people or sometimes dragged to the vehicle capable of transporting them. It is obvious that a wild boar opened from the jaws to the pelvis and dragged for hundreds of meters will tend to be contaminated, although these contaminations are to be considered secondary for the preservation of the meat, compared to contamination by the intestinal contents. In an attempt to investigate the effect of delayed refrigeration on wild boar carcass contamination, the aim of this work was to determine a correlation between several hunting and logistic parameters (age, sex, animal weight, shooting distance, number of shots, weather and temperature and time from shot to refrigeration and to analysis) and bacterial contamination of the carcass. The correlation coefficient, r, was found to be 0.038 for the eviscerated body weight ( p < 0.05), 0.091 for the external temperature on the day of hunting ( p < 0.05), 0.027 for the time from shot to refrigeration ( p = 0.081), 0.038 for the time from refrigeration to analysis ( p < 0.05) and 0.043 for the time from shot to analysis ( p < 0.05). These results stand for a negative correlation between the bacterial population and eviscerated carcass weight and between the bacterial population and external temperature and for a positive correlation between the time from shot to analysis and from refrigeration to analysis. No association was demonstrated between the bacterial population and the time from shot to refrigeration.

In this research communication we report on the diversity of yeast and mould species in 69 samples of milk and different dairy products from three plants located in Umbria, central Italy. Isolates were characterised both macroscopically and microscopically and then identified by PCR and genome sequencing of the ITS region and the D1-D2 domain of the large-subunit rRNA gene for filamentous fungi and yeasts, respectively. Out of the 69 samples analysed, 51 (73.9%) tested positive for the presence of yeasts, whereas moulds were detected in 25 (36.2%) samples. A total of 9 yeast species belonging to 8 different genera and 13 mould species belonging to 6 different genera were isolated. The most common genera isolated were Debaryomyces and Kluyveromyces among the yeasts and Penicillium and Galactomyces among the moulds. Microbiota play a key role in the formation of flavour, aroma, texture and appearance of dairy products. This complex microbial ecosystem includes both cultured and external bacteria, yeasts and moulds. Some of them have an important role in the production of cheeses, whereas others are responsible for dairy product spoilage, resulting in significant food waste and economic losses. Some species can produce mycotoxins, representing a potential hazard for the consumer's safety. This study provides interesting information on the diversity of fungi species in dairy products from central Italy that can be of major importance to identify these products and to develop adequate strategies for fungal spoilage control and consumer safety.

This study describes a simple method for the large-scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites. T. gondii tachyzoites were obtained from infected human foreskin fibroblasts (HFFs) and peritoneal exudates of mice, while tissue cysts containing bradyzoites were collected from chronically infected mice. Harvested cells and brain tissues were incubated in Hanks balanced salt solution (HBSS), containing 0.25% trypsin and 0.5% taurodeoxycholic acid (TDC) for 5 min. Subsequent washes in phosphate buffered saline (PBS) were conducted, and the cell viability of the preparations was good, as determined by flow cytometry and ability to reinfect HFF cells and propagate in mice. The purification procedure allowed for a rapid preparation of pure T. gondii tachyzoites and bradyzoites in sufficient quantity that can be used for downstream procedures. The advantage of the new method is that it is convenient and inexpensive., (© 2020 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Ground beef contamination with Escherichia coli is usually a result of carcass faecal contamination during the slaughter process. Carcasses are contaminated when they come into contact with soiled hides or intestinal leakage content during dressing and the evisceration processes. A more recent and compelling hypothesis is that, when lymph nodes are present in manufacturing beef trimmings, they can be a potential source of Enterobacteriaceae contamination of ground beef. The aim of this study was to investigate the occurrence of E. coli in lymph nodes from beef carcasses used for ground meat production, in six slaughter plants situated in central Italy A total of 597 subiliac (precrural) lymph nodes were obtained from 597 cattle carcasses and screened for E. coli by culture. Furthermore, E. coli isolates (one per positive carcass) were tested for stx1, stx2 eaeA and hlyA genes that are commonly used to identify and characterise shiga toxin-producing E. coli (STEC). In addition, the E. coli isolates were profiled for antimicrobial susceptibility. A proportion of 34.2% (204/597) carcasses were positive for E. coli. PCR revealed that 29% (59/204) of E. coli possessed stx1 or stx2 which corresponded to 9.9% of the cattle sampled. Moreover, a combination of stx1 or stx2 and eaeA was found in in 4 isolates (2% among E. coli positive samples and 1% among cattle sampled) and a combination of stx1 or stx2 and eaeA and hly in 1 isolate (0.5% and 0.2%). More than 95% of isolates were susceptible to gentamicin, ceftriaxone, cyprofloxacin and cefotaxime while high rates of resistance were recorded for cephalotin, ampicillin, tetracycline, tripe sulfa and streptomycin. The multivariate analysis identified "age" as the factor most closely related to E. coli positivity (either generic E. coli or STEC) in bovine lymph nodes. In conclusion, subiliac lymph nodes represent a source of E. coli for ground beef. These results are of major importance for risk assessment and improving good manufacturing practices during animal slaughter and ground meat production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Community-based nutritional intervention to improve the practice of dietary diversity and child nutrition by community health workers (CHWs) involving Nyumba Kumi as small neighborhood units (SNUs) in communities has not yet been explored. This study was conducted in two villages in rural Kenya between 2018 and 2019. In total, 662 participants (control vs. intervention: n = 339 vs. n = 323) were recruited. The intervention group received education on maternal and child nutrition and follow-up consultations. The custom-tailored educational guidelines were made based on Infant and Young Child Feeding and the mother and child health booklet. The educational effects on household caregivers' feeding practice attitude and child nutritional status were analyzed using multiple linear regression. After the intervention, a total of 368 household caregivers (187 vs. 181) and 180 children (113 vs. 67) were analyzed separately. Between the groups, no significant difference was found in their background characteristics. This study successfully improved the dietary diversity score (β = 0.54; p < 0.01) and attitude score (β = 0.29; p < 0.01). The results revealed that the interventions using CHWs and SNUs were useful to improve dietary diversity and caregivers' attitudes toward recommended feeding. This research has the potential to be successfully applied in other regions where child undernutrition remains.

This study investigated occurrence and antimicrobial resistance profiles of Campylobacter spp. isolates in beef cattle on five cow-calf operations in South Africa. A total of 537 fecal samples from adult beef cattle ( n  = 435) and rectal swabs from calves ( n  = 102) were screened for Campylobacter jejuni , Campylobacter coli , and Campylobacter upsaliensis by culture and polymerase chain reaction. Furthermore, 86 Campylobacter spp. isolates including 46 C. jejuni , 24 C. coli , and 16 C. upsaliensis were tested for antimicrobial resistance against a panel of 9 antimicrobials. Overall, Campylobacter spp. was detected in 29.7% of cattle. Among the 158 Campylobacter spp.-positive cattle, 61.8% carried C. jejuni , 25% carried C. coli , and 10% carried C. upsaliensis . Five animals (3.1%) had mixed infections: three cows carried C. jejuni and C. coli concurrently, one cow had both C. jejuni and C. upsaliensis , and one cow harbored C. coli and C. upsaliensis . Antimicrobial resistance profiling among 86 Campylobacter spp. isolates revealed that 52.3% of the isolates were resistant to one or more antimicrobials. Antimicrobial resistance was observed in 46.7% of C. jejuni isolates, 35.6% of C. coli , and 17.8% of C. upsaliensis . Thirty-six percent of isolates were resistant to clindamycin, 19.7% to nalidixic acid, 18.6% to tetracycline, and 17.4% to erythromycin. Lower resistance rates were recorded for azithromycin (8.1%), florfenicol (3.4%), gentamicin (4.8%), and telithromycin and ciprofloxacin (5.8%). Multidrug resistance (MDR) was observed in 32.5% of isolates. Significantly higher levels of MDR were detected among C. jejuni (36.9%) and C. coli (33.3%) isolates in comparison to C. upsaliensis (18.7%). Two main multiresistance patterns were detected: nalidixic acid/clindamycin (17.8%) and tetracycline/clindamycin (14.2%). To the best of our knowledge, this is the first study which has shown that beef cattle on cow-calf operations in South Africa constitute an important reservoir and a potential source of clinically relevant and antimicrobial resistant Campylobacter spp. strains.

Abstract: The presence of lactic acid bacteria can be detrimental when the abundant growth of slime-producing strains (Lactobacillus spp. and Leuconostoc spp.) causes spoilage of meat products. Two strains of lactic acid bacteria were isolated from vacuum-packed cooked hams that had been withdrawn from the market for the so-called ropy slime defect and identified as Leuconostoc mesenteroides. In an attempt to define the behavior of ropy slime-producing bacteria, two strains of L. mesenteroides were incubated in de Man Rogosa Sharpe broth at different storage temperatures and conditions of thermal abuse (4, 12, 20, 30, 37, and 44°C). Both strains showed a lack of growth at 44°C, a good level of development at 30 and 37°C, and evident growth ability at low temperatures, with a long stationary phase. In particular, the bacterial concentration at 4°C was >105 CFU mL-1 after more than 120 days of incubation. This study demonstrates that the refrigeration temperature for cooked meat products does not constitute a hurdle for ropy slime producers and their subsequent ability to spoil., (Copyright ©, International Association for Food Protection.)

Background: Non-communicable diseases (NCDs) are increasing in Kenya, where HIV/AIDS remains a leading cause of death; however, few studies have investigated obesity and hypertension among adults with HIV infection. We conducted a cross-sectional study in Homa Bay, Western Kenya, during 2015 to determine the prevalence of overweight/obesity and hypertension among HIV-infected adults and to identify their risk factors., Results: Anthropometric measurements and a structured questionnaire were administered to adults with HIV infection receiving care at Mbita Sub-county Hospital. A total of 251 HIV-positive individuals were enrolled. More women were overweight (17.2%) and obese (3.6%) than underweight (8.3%). The prevalence of abdominal obesity was high in women (62.7%), especially those aged 30-39 years. The prevalence of hypertension was 9.8% and 11.8% in men and women, respectively. Male participants tended to develop hypertension at an early age. Multivariate analysis showed that female sex was significantly associated with abdominal obesity. Regarding clinical factors, we identified an association between overweight and a history of opportunistic infections, as well as between hypertension and World Health Organization clinical stage. Sixty percent of HIV-infected participants assumed that a very thin body size indicated HIV infection., Conclusions: The main findings of this study include a greater prevalence of overweight than underweight as well as a high prevalence of abdominal obesity among women. Social perception toward body size among people with HIV infection might remain problematic. Individuals living with HIV in Kenya should receive preventive intervention for overweight and abdominal obesity, with consideration of relevant social and cultural aspects., Competing Interests: Competing interestsNo potential conflict of interest was reported by the authors., (© The Author(s) 2020.)

The use antimicrobials for therapeutic and metaphylactic purpose in humans and agriculture exerts selective pressure on animal and environmental microbiota resulting in the survival and spread of antimicrobial resistance genes among bacteria and subsequent development of resistance in bacteria. Previous studies have shown that honey bees' microbiota (Apis mellifera) can accumulate antimicrobial resistance genes in their microbiome and act as collectors and disseminators of resistance genes. The aim of this study was to investigate to what extent honey bees act as reservoir of select antimicrobial resistance genes. This study was conducted on 35 groups of bees. Bees were collected from 35 sites in Umbria, Italy. PCR was used to screen pooled ground bees' specimens for genes that code for resistance against antimicrobials that are commonly used in humans and in veterinary medicine including aminoglycosides (aph), beta-lactams (blaZ), tetracycline (tetM) and sulphonamides (sul1 and sul2). Twenty-four samples out of 35 (68.57%) were positive for at least one antimicrobial resistance gene. Two samples were positive for the aph, 5.71%; eight for blaZ, 22.86%; three for tetM, 8.57%; ten for sul1, 28.57% and eighteen for sul2, 51.43%. Positivity to more than one antimicrobial resistance gene was observed in nine samples, 25.71%. The multivariate analysis identified "presence of farms nearby" as the factor most closely related to PCR positivity. Honey bees (Apis mellifera) from Umbria, Italy, carry antimicrobial resistance genes and can be used as indicators of the presence of resistance genes in the environment.

The early diagnosis of mastitis is an essential factor for the prompt detection of the animal for further actions. In fact, if not culled, infected cows must be segregated from the milking herd and milked last, or milked with separate milking units. Besides microbiological analysis, the somatic cell count (SCC) commonly used as predictor of intramammary infection, frequently lead to a misclassification of milk samples. To overcome these limitations, more specific biomarkers are continuously evaluated. The total amino acid content increases significantly in mastitic milk compared to normal milk. S. aureus requires branched-chain amino acids (BCAAs-isoleucine, leucine, and valine) for protein synthesis, branched-chain fatty acids synthesis, and environmental adaptation by responding to their availability via transcriptional regulators. The increase of BCAAs in composite milk has been postulated to be linked to mammary infection by S. aureus . The aim of this work is to demonstrate, by a direct ion-pairing reversed-phase method, based on the use of the evaporative light-scattering detector (IP-RP-HPLC-ELSD), applied to 65 composite cow milk samples, a correlation between the concentration of isoleucine and leucine, and S. aureus load. The correlation coefficient, r, was found to be 0.102 for SCC ( p = 0.096), 0.622 for isoleucine ( p < 0.0001), 0.586 for leucine ( p < 0.0001), 0.013 for valine ( p = 0.381), and 0.07 for tyrosine ( p = 0.034), standing for a positive correlation between S. aureus and isoleucine and leucine concentration. The link between the content of BCAAs, isoleucine and leucine, and udder infection by S. aureus demonstrated with our study has an important clinical value for the rapid diagnosis of S. aureus mastitis in cows.

In this study, 140 cattle STEC isolates belonging to serogroups O157, O26, O145, O121, O103 and O45 were characterized for 38 virulence-associated genes, antimicrobial resistance profiles and genotyped by PFGE. The majority of isolates carried both stx1 and stx2 concurrently, stx2c, and stx2d; plasmid-encoded genes ehxA, espP, subA and saa but lacked katP and etpD and eaeA. Possession of eaeA was significantly associated with the presence of nle genes, katP, etpD, ureC and terC. However, saa and subA, stx1c and stx1d were only detected in eaeA negative isolates. A complete OI-122 and most non-LEE effector genes were detected in only two eaeA positive serotypes, including STEC O157:H7 and O103:H2. The eaeA gene was detected in STEC serotypes that are commonly implicated in severe humans disease and outbreaks including STEC O157:H7, STEC O145:H28 and O103:H2. PFGE revealed that the isolates were highly diverse with very low rates of antimicrobial resistance. In conclusion, only a small number of cattle STEC serotypes that possessed eaeA, had the highest number of virulence-associated genes, indicative of their high virulence. Further characterization of STEC O157:H7, STEC O145:H28 and O103:H2 using whole genome sequencing will be needed to fully understand their virulence potential for humans.

Objectives: Stunting is a significant cause of poor cognitive performance and lower school achievement. Stunting is observed among pre-school children in several areas in Africa; however, not all children are affected, and children with and without stunting are seen in the same communities. Therefore, this study aimed to identify nutritional and other factors that prevent stunting that may exist in local communities., Study Design: This is a prospective cohort study., Methods: Data were extracted from the Health and Demographic Surveillance System conducted in Kwale County, Kenya. The cohort consisted of all households with children less than five years old, within a radius of 2.2 km from a local health centre. A dietary pattern (DP) survey with a semi-quantitative food frequency questionnaire was conducted on caretakers of children who were voluntary participated from the cohort between June 2012 and August 2012. Using cluster analysis, the children were assigned to a DP group. Logistic regression analysis was applied to calculate the adjusted odds ratios (aORs) of DPs for stunting controlling for other factors., Results: In total, 402 children were included in the analysis. By cluster analysis, three DPs were identified: protein-rich DP; traditional DP; and traditional DP complemented by breastfeeding. The aOR of a child becoming stunted from a normal height during the study period among children who received a traditional DP compared with those who had a protein-rich DP was 2.78 (95% confidence interval [CI]: 1.02-7.55). However, the aOR for children who were already stunted at the start of the study and had a traditional DP was 1.49 (95% CI: 0.82-2.72). Increased aORs of stunting were observed among children aged over 12 months compared with children aged 6-11 months, and the effects of DPs were modified by age in months from 12 to 35 months; however, the effects were near the null value for children over 36 months of age, although these were not statistically significant., Conclusions: We found that the traditional DP showed a higher risk for stunting compared with the protein-rich DP, and the most vulnerable age range for stunting was between 12 and 35 months. Interventions to prevent stunting should focus on providing 12- to 35-month-old children with locally available, protein-rich foods., (Copyright © 2019 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Shiga toxin-producing Escherichia coli (STEC) isolates (N = 38) that were incriminated in human disease from 2006 to 2013 in South Africa were characterized by serotype, virulence-associated genes, antimicrobial resistance and pulsed-field gel electrophoresis (PFGE). The isolates belonged to 11 O:H serotypes. STEC O26:H11 (24%) was the most frequent serotype associated with human disease, followed by O111:H8 (16%), O157:H7 (13%) and O117:H7 (13%). The majority of isolates were positive for key virulence-associated genes including stx1 (84%), eaeA (61%), ehxA (68.4%) and espP (55%), but lacked stx2 (29%), katP (42%), etpD (16%), saa (16%) and subA (3%). stx2 positive isolates carried stx2c (26%) and/or stx2d (26%) subtypes . All pathogenicity island encoded virulence marker genes were detected in all (100%) isolates except nleA (47%), nleC (84%) and nleD (76%). Multidrug resistance was observed in 89% of isolates. PFGE revealed 34 profiles with eight distinct clusters that shared ≥80% intra-serotype similarity, regardless of the year of isolation. In conclusion, STEC isolates that were implicated in human disease between 2006 and 2013 in South Africa were mainly non-O157 strains which possessed virulence genes and markers commonly associated with STEC strains that have been incriminated in mild to severe human disease worldwide. Improved STEC monitoring and surveillance programs are needed in South Africa to control and prevent STEC disease in humans., Competing Interests: The authors declare no conflict of interest.

Reports on the occurrence of Campylobacter spp. in dogs in South Africa are non-existent. This study investigated the prevalence of Campylobacter spp. in 481 dogs visiting four rural community veterinary clinics in South Africa. Dogs were screened for Campylobacter spp. by culture and polymerase chain reaction (PCR), and logistic regression analysis was performed to assess the association between sex, clinic, breed and age and the occurrence of Campylobacter spp. in dogs. The prevalence of Campylobacter spp. was 41.50% (95% confidence interval [CI], 37.39% - 46.04%). Campylobacter jejuni, C. upsaliensis and C. coli were detected in 29.31% (95% CI, 25.42% - 33.54%), 13.10% (95% CI, 10.37% - 16.42%) and 5.41% (95% CI, 3.71% - 7.82%) of dogs, respectively. Dogs carrying more than one species of Campylobacter spp. accounted for 6.23% (95% CI, 4.40% - 8.78%). Campylobacter upsaliensis and C. jejuni were detected in 3.74% (95% CI, 2.37% - 5.86%), whereas C. coli and C. jejuni were found in 2.49% (95% CI, 1.42% - 4.34%) of dogs. Age and clinic were the risk factors significantly associated with Campylobacter spp. occurrence, while age, breed and clinic were predictors of C. jejuni carriage. Furthermore, age was the only risk factor associated with a higher likelihood of carrying C. upsaliensis. The prevalence of Campylobacter spp. C. jejuni and C. upsaliensis increased significantly as dogs grew older. In addition, the odds of carrying Campylobacter spp. were higher in the Staffordshire bull terrier breed compared to crossbreed dogs. In conclusion, this study shows that dogs visiting rural community veterinary clinics in South Africa are reservoirs of Campylobacter spp. and may be potential sources of Campylobacter spp. for humans living in close proximity of the dog populations under study.

Possible contamination by Staphylococcus aureus of the production environment and of the meat of a canned meat production factory was analysed. A total of 108 samples were taken from nine critical control points, 13 of them were positive for S . aureus . None of the isolates produced enterotoxins. To determine how much time can elapse between can seaming and sterilisation in the autoclave without any risk of enterotoxin production by S . aureus , the growth and enterotoxin production of three enterotoxin A producing strains of S. aureus (one ATCC strain and two field strains) in canned meat before sterilisation was investigated at three different temperatures (37, 20 and 10 °C). Two types of meat were used, one with and one without sodium nitrite. In the canned products, the spiked bacteria spread throughout the meat and reached high levels. Enterotoxin production was shown to start 10 hours after incubation at 37 °C and after 48 h after incubation at 20 °C; the production of enterotoxin was always detected in the transition between the exponential and the stationary growth phase. At 10 °C, the enterotoxin was never detected. The statistical analysis of the data showed that the difference between the two different types of meat was not statistically significant ( p value > 0.05). Since it is well known that following heat treatment, staphylococcal enterotoxins, although still active (in in vivo assays), can be undetectable (loss of serological recognition) depending on the food matrix and pH, it is quite difficult to foresee the impact of heat treatment on enterotoxin activity. Therefore, although the bacteria are eliminated, the toxins may remain and cause food poisoning. The significance of the results of this study towards implementing good manufacturing practices and hazard analysis critical control points in a canned meat factory are discussed with reference to the management of pre-retorting steps after seaming.

Avian influenza virus subtype H9N2 is identified in chickens with respiratory disease while Bacillus cereus (B. cereus) has been frequently isolated from chicken feed in China. However, the roles of co-infection with these two pathogens remain unclear. In the present study, SPF chicks were intragastrically administered with 10 8  CFU/mL of B. cereus for 7 days and then inoculated intranasally with 100 EID 50 of H9N2 three days later. Alternatively, chickens were initially inoculated with H9N2 and then with B. cereus for one week. Post administration, typical respiratory distress persisted for 5 days in both co-infection groups. Gizzard erosions developed in the groups B. cereus/H9N2 and B. cereus group on 7 th day while in group H9N2/B. cereus on 14 th day. More importantly, both air-sac lesions and lung damage increased significantly in the co-infection group. Significant inflammatory changes were observed in the B. cereus group from day 7 to day 21. Moreover, higher loads of H9N2 virus were found in the co-infected groups than in the H9N2 group. Newcastle Disease Virus (NDV) specific antibodies were decreased significantly in the H9N2/B. cereus group compared to the B. cereus and the B. cereus/H9N2 groups. Nonspecific IgA titers were reduced significantly in the B. cereus group and the H9N2/B. cereus group compared to the control group. In addition to this, lower lymphocyte proliferation was found in the con-infection groups and the H9N2 group. Hence, feed-borne B. cereus contamination potentially exacerbates gizzard ulceration and aggravates H9N2-induced respiratory distress by inhibiting antibody-mediated immunity and pathogen clearance. Thus controlling the B. cereus contamination in poultry feed is immediately needed.

Staphylococcus aureus is not only a common cause of bovine mastitis, but also an agent of food poisoning in humans. In an attempt to determine whether staphylococci causing bovine mastitis could also cause food poisoning, 60 isolates of presumed S. aureus were isolated in the period between March and August 2017 from 3,384 routine, composite, quarter milk samples of individual cows raised on 12 dairy farms in central Italy. Seventeen out of 60 isolates were confirmed as S. aureus after coagulase, thermonuclease, and biochemical tests. These isolates were analyzed by PCR for the presence of the nuc, sea, seb, sec, sed, and see genes. The positive isolates were nuc, 100% (17); sea, 35.29% (6); seb, 5.88% (1); sec, 5.88% (1); sed, 29.41% (5); and see, 47.06% (8). The isolates were also tested with 2 enzyme immunoassay diagnostic kits, one for the screening detection of the production of staphylococcal enterotoxins (SEA, SEB, SEC, SED, SEE) and one for the detection of specific enterotoxin produced by each isolate. Seven out of 17 (41.18%) were enterotoxin producers: 7 produced SEA (41.18%), 1 SEB (5.88%), 1 SEC (5.88%), 5 SED (29.41%), and 6 SEE (35.29%). To further characterize the isolates, they were analyzed by the Kirby Bauer test for susceptibility to 13 antimicrobials (ampicillin, ciprofloxacin, kanamycin, tetracycline, gentamicin, methicillin, nalidixic acid, erythromycin, amoxicillin/clavulanic acid, streptomycin, vancomycin, neomycin, and enrofloxacin), and we detected resistance to ampicillin (52.94%), nalidixic acid (70.59%), erythromycin (5.88%), and amoxicillin/clavulanic acid (17.65%). The isolates were sensitive to the main classes of antimicrobials used for the treatment of bovine subclinical mastitis. The presence of enterotoxin-producing isolates of S. aureus in bovine milk means that a temperature abuse or a breakdown in the thermal treatment of the milk could present a food safety risk, particularly if all enterotoxigenic isolates could potentially produce SEA in milk., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Cattle are a major reservoir of Shiga toxin-producing Escherichia coli. This study investigated the occurrence of seven major STEC serogroups including O157, O145, O103, O121, O111, O45 and O26 among 578 STEC isolates previously recovered from 559 cattle. The isolates were characterized for serotype and major virulence genes. Polymerase chain reaction revealed that 41.7% (241/578) of isolates belonged to STEC O157, O145, O103, O121, O45 and O26, and 33 distinct serotypes. The 241 isolates corresponded to 16.5% (92/559) of cattle that were STEC positive. The prevalence of cattle that tested positive for at least one of the six serogroups across the five farms was variable ranging from 2.9% to 43.4%. Occurrence rates for individual serogroups were as follows: STEC O26 was found in 10.2% (57/559); O45 in 2.9% (16/559); O145 in 2.5% (14/559); O157 in 1.4% (8/559); O121 in 1.1% (6/559); and O103 in 0.4% (2/559). The following proportions of virulence genes were observed: stx1, 69.3% (167/241); stx2, 96.3% (232/241); eaeA, 7.1% (17/241); ehxA, 92.5% (223/241); and both stx1 and stx2, 62.2% (150/241) of isolates. These findings are evidence that cattle in South Africa carry STEC that belong to six major STEC serogroups commonly incriminated in human disease. However, only a subset of serotypes associated with these serogroups were clinically relevant in human disease. Most STEC isolates carried stx1, stx2 and ehxA but lacked eaeA, a major STEC virulence factor in human disease., (© 2018 Blackwell Verlag GmbH.)

This study evaluated the effect of a novel formulation for starter culture associated with specific ripening conditions (NoNit™ technology) vs. a commercial» starter on the fate of selected pathogens and hygiene indicators during the fermentation and ripening of experimentally spiked salame nostrano (Italian dry sausage). Selected strains of Staphylococcus aureus 27R, Escherichia coli CSH26 K 12, Listeria innocua ATCC 33090 and Salmonella Derby 27 were inoculated into salami batter and challenged with two formulations of starter cultures (a commercial formulation and the NoNit™ formulation, consisting of Lactococcus lactis ssp. lactis, strain 340; L. lactis ssp. lactis, strain 16; Lactobacillus casei ssp. casei, strain 208 and Enterococcus faecium strain 614) with ripening at a low temperature. The proposed technology (NoNit™) performed better than the commercial formulation and limited the growth of spiked Escherichia coli, Staphylococcus aureus (including the production of enterotoxin), Salmonella Derby and Listeria innocua, yet maintained the basic product appearance and texture., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

We have set up an ex vivo ovine ruminal model, which can mimic the multicellular process to explore the early steps in Salmonella typhimurium lipopolysaccharide (LPS) stimulation using RNA-seq technology. Ovine ruminal explants were collected for histological and transcriptional analysis and supernatants collected to quantitate lactate dehydrogenase (LDH) enzymes. A total of 8 and 523 genes were significantly over-expressed between LPS-treated and control tissues at 6 and 12 h, respectively. However, six and seven hundred and thirteen genes were substantially repressed between the aforementioned tissues, correspondingly. Key genes up-regulated in response to the addition of LPS were tumor necrosis factor (TNF), interlukin (IL)-1 beta(b), IL-6, IL-8, IL-17B, IL-19, MMP-1, MMP-3, and integrin alpha 2 (ITGA8, 9). This study shows for the first time that galectin-1 is up-regulated in an ex vivo ruminal segment model exposed to bacterial lipopolysaccharide following 6 h of incubation. The ruminal segment model has been shown to be a suitable tool to study the bacterial lipopolysaccharide effects on the ovine ruminal tissues prior to in vivo assessment.