Evaluation of circulating microRNA profiles in blood as potential candidate biomarkers in a subacute ruminal acidosis cow model - a pilot study.

BACKGROUND: Subacute ruminal acidosis (SARA) is a metabolic disorder often observed in high-yielding dairy cows, that are fed diets high in concentrates. We hypothesized that circulating miRNAs in blood of cows could serve as potential candidate biomarkers to detect animals with metabolic dysbalances such as SARA. MicroRNAs (miRNAs) are a class of small non-coding RNAs, serving as regulators of a plethora of molecular processes. To test our hypothesis, we performed a pilot study with non-lactating Holstein-Friesian cows fed a forage diet (FD; 0% concentrate, n = 4) or a high-grain diet (HG; 65% concentrate, n = 4) to induce SARA. Comprehensive profiling of miRNA expression in plasma and leucocytes were performed by next generation sequencing (NGS). The success of our model to induce SARA was evaluated based on ruminal pH and was evidenced by increased time spent with a pH threshold of 5.8 for an average period of 320 min/d. RESULTS: A total of 520 and 730 miRNAs were found in plasma and leucocytes, respectively. From these, 498 miRNAs were shared by both plasma and leucocytes, with 22 miRNAs expressed exclusively in plasma and 232 miRNAs expressed exclusively in leucocytes. Differential expression analysis revealed 10 miRNAs that were up-regulated and 2 that were down-regulated in plasma of cows when fed the HG diet. A total of 63 circulating miRNAs were detected exclusively in the plasma of cows with SARA, indicating that these animals exhibited a higher number and diversity of circulating miRNAs. Considering the total read counts of miRNAs expressed when fed the HG diet, differentially expressed miRNAs ( log2 fold change) and known function, we have identified bta-miR-11982, bta-miR-1388-5p, bta-miR-12034, bta-miR-2285u, and bta-miR-30b-3p as potential candidates for SARA-biomarker in cows by NGS. These were further subjected to validation using small RNA RT-qPCR, confirming the promising role of bta-miR-30b-3p and bta-miR-2285. CONCLUSION: Our data demonstrate that dietary change impacts the release and expression of miRNAs in systemic circulation, which may modulate post-transcriptional gene expression in cows undergoing SARA. Particularly, bta-miR-30b-3p and bta-miR-2285 might serve as promising candidate biomarker predictive for SARA and should be further validated in larger cohorts.

Methicillin-resistant Staphylococcus aureus (MRSA) isolated from raw milk and nasal swabs of small ruminants in Abeokuta, Nigeria.

Methicillin-resistant Staphylococcus aureus (MRSA), an important widespread cause of severe infection in both humans and animals, is a significant pathogen of public health concern. This study examined the presence of MRSA in 400 samples comprising 200 raw milks (145 from goat and 55 from sheep) and 200 nasal swabs (145 from goats and 55 from sheep) collected from ten different locations in Abeokuta, Nigeria. Samples were examined using standard bacteriological methods for the isolation and identification of Staphylococcus aureus and culture on oxacillin (6 µg/ml) and cefoxitin (2 µg/ml) selective media for the detection of MRSA. Suspected MRSA isolates were confirmed by latex agglutination test for the detection of penicillin-binding protein 2a (PBP2a). Antibiotic susceptibility testing was determined by Kirby Bauer disc diffusion method. Staphylococcus aureus was detected in 72 (18%) of 400 samples of which 52 (13%) were confirmed as MRSA. Methicillin-resistant S. aureus was detected in raw milk (37 of 200; 18.5%) and nasal swab (15 of 200; 7.5%). There was no significance difference (p > 0.05) in the prevalence of MRSA in sheep (37.7%) and goat (23.4%). The MRSA isolates showed resistance to ampicillin (100%), cloxacillin (100%), sulphamethoxazole-trimethoprim (100%), amoxicillin-clavulanate (84.6%), ceftriaxone (75%), cefuroxime (69.2%), erythromycin (65.4%), streptomycin (38.5%), ciprofloxacin (23.1%), pefloxacin (21.2%) and gentamicin (17.3%). The presence of multidrug-resistant MRSA in small ruminants reared in Abeokuta metropolis may be due to regular use of antibiotics and unhygienic practices by farmers. This in turn constitutes a potential public health risk to the owners, consumers of small ruminant products and the general populace.

Multidrug-resistant Salmonellae isolated in Japanese quails reared in Abeokuta, Nigeria.

Salmonellosis is a major bacterial disease causing huge economic losses in the poultry industry worldwide. This study was carried out to determine the period prevalence and antimicrobial susceptibility of Salmonella enterica in Japanese quails in Abeokuta, Nigeria. Four hundred cloacal swabs of quail birds were collected from 4 locations within Abeokuta. Salmonella was isolated from the samples using conventional methods for selective isolation of Salmonella and biochemical identification. Isolates were confirmed by polymerase chain reaction assays for the amplification and detection of Salmonella-associated virulence genes (invA and stn) using specific primers. Antimicrobial susceptibility testing was done using the Kirby-Bauer disk diffusion method. In all, Salmonella was isolated from 14 (3.5%) cloacal swabs. All 14 isolates possessed invA and stn genes. The Salmonella isolates showed resistance to tetracycline (100%), doxycycline (100%), ampicillin (100%), sulphamethoxazole (92.9%), nalidixic acid (85.8%), ceftazidime (78.6%), neomycin (64.3%), streptomycin (50%) and gentamycin (28.6%) but all the isolates were susceptible to ciprofloxacin. The isolates were resistant to at least three antimicrobials indicating multidrug resistance. The results concluded that Japanese quails harbour multidrug-resistant Salmonella which could be transmitted to humans through consumption of contaminated food or by direct and indirect contact with the carrier birds. Antimicrobial resistance could be due to overdependence on antimicrobials. Ciprofloxacin could be considered in the treatment of zoonotic Salmonellosis in humans.

Iron status and benefit of the use of parenteral iron therapy in pre-dialysis Chronic Kidney disease patients.

AIMS AND OBJECTIVES: Anaemia is a major cardiovascular risk factor in chronic kidney disease (CKD). Treatment and correction of anaemia leads to improvement of cardiovascular status and quality of life of patients with CKD. This interventional open labeled randomised controlled study comparing the effect of intravenous and oral iron therapy in improving red cell indices and iron status in anaemic pre-dialysis Chronic Kidney Disease was carried out to determine iron status in anaemic pre-dialysis CKD patients to assess the benefit of parenteral iron supplementation as against empirical oral iron in CKD patients with iron deficiency. PATIENTS AND METHODS: Sixty consecutive pre-dialysis chronic kidney disease patients attending the renal clinic over a six month period were screened. Forty- one subjects (68.3%) were found to be anaemic and were subsequently studied. RESULTS: The ages of the patients ranged between 19 and 71 years with a mean age of 39 years. The mean serum creatinine and mean creatinine clearance were 201.80 (70.25)?mol/L and 37.90 (± 12.17)ml/minute respectively. The haematocrit concentration was found to correlate inversely with the level of serum creatinine and 56.1% of the anaemic patients had iron deficiency. The mean PCV rise in the intravenous iron group was 2.42 (± 1.98)% and this was statistically significant (p=0.002) while the mean PCV difference was 0.909 (± 0.94)% in the oral iron group. Intravenously administered iron alone permitted anaemia correction in about one-third of these patients without any life threatening adverse drug event. CONCLUSION: Anaemia is very common in the pre-dialysis CKD population and the prevalence of iron deficiency is high. Intravenous iron supplementation is an effective and safe treatment for the anaemia in the pre-dialysis CKD patients. Response to oral iron was poor.

Evolution of Salmonella nomenclature: a critical note.

Salmonellae are widely distributed but nomenclaturally controversial pathogens of both humans and animals. Despite elaborate studies, much still remain to be discovered about these organisms. Although Salmonella nomenclature has proved to be rather complex, in 2005, Salmonella enterica finally gained official approval as the type species of the genus Salmonella. In addition, one other species has been approved and recognised in the genus Salmonella, namely, Salmonella bongori. New serovars (serotypes) are continually being discovered each year and reported in the journal Research in Microbiology. Salmonella serovars and their antigenic formulae are listed in the White-Kauffmann-Le Minor scheme and updated by the World Health Organization's Collaborating Centre for Reference and Research on Salmonella at the Pasteur Institute, Paris, France.

Potentially zoonotic shiga toxin-producing Escherichia coli serogroups in the faeces and meat of food-producing animals in Ibadan, Nigeria.

Shiga toxin-producing Escherichia coli (STEC) are major food-borne pathogens associated with gastroenteritis and sometimes fatal haemolytic uraemic syndrome complication. Farm animals are asymptomatic carriers of STEC and contaminated meat is an important vehicle for zoonotic transmission from animals to humans. This study investigated the presence, virulence traits and antimicrobial susceptibility of seven potentially human pathogenic STEC serogroups (O157, O26, O91, O103, O111, O128 and O145) in the faeces and meat of food-producing animals in Ibadan, Nigeria. One hundred and fifty-four (7.3%) of 2133 samples were positive for STEC serogroups. The pathogens were detected in the faeces of cattle (15.2%), sheep (10.7%), goats (7.5%) and pigs (5.6%) as well as in beef (3.8%), goat-meat (1.7%) and pork (4.0%). All seven investigated STEC serogroups were found in cattle, all except O145 were found in sheep, three serogroups (O157, O26 and O111) were found in goats and three (O157, O111 and O128) in pigs. The rate of detection of each of the serogroups in all 2133 samples was: O157 (5.0%), O26 (0.2%), O91 (0.3%), O103 (0.3%), O111 (1.0%), O128 (0.2%) and O145 (0.1%). Of all 154 isolates, 11.0% had shiga toxin type 1 gene (stx(1)), 25.3% had stx(2) and 41.6% had stx(1)/stx(2); intimin gene (eaeA) was detected in 56.5% and enterohaemolysin gene (hlyA) in 75.3%. Among the O157 isolates, 24.5% were negative for stx genes but positive for eaeA and/or hlyA while 7.6% were negative for all four virulence genes. Fourteen different combinations of virulence genes were encountered but stx(1)/stx(2)/eaeA/hlyA combination was the most predominant. The percentage resistance of the isolates to the tested antimicrobial agents was: ampicillin (82.5%), chloramphenicol (42.9%), ciprofloxacin (22.1%), enrofloxacin (25.3%), nalidixic acid (37.7%), neomycin (24.0%), norfloxacin (20.8%), streptomycin (50.7%) and tetracycline (75.3%). One hundred and forty-eight (96.1%) of all 154 isolates were resistant to at least one of the tested antimicrobial agents while 69.5% were categorised as multi-drug resistant. Potentially pathogenic multi-drug resistant STEC isolates were recovered from the meat production chain in Nigeria. Unhygienic practices that predominate during slaughter and processing were observed to have contributed to faecal contamination and presence of STEC in meat.

Acute renal failure (ARF) in developing countries: which factors actually influence survival.

Acute renal failure, a relatively common clinical condition, is still associated with a high mortality rate in both developed and developing countries despite the differences in the epidemiology, clinical characteristics and therapeutic modalities offered to affected patients. The various modalities of renal replacement therapy are still largely inaccessible, unaffordable and unavailable in most parts of sub-Saharan Africa, hence the need to judiciously utilise available resources. Consequently we studied patients with acute renal failure to critically appraise the factors that influence survival and determine the usefulness or otherwise of available renal replacement therapies (Acute HD and Acute PD). A total of 46 (34 (73.9%) males and 12 (26.1%) females) patients satisfied the inclusion criteria. Their ages ranged between 15 and 76 years (mean +/- SD; 38.2 +/-16.3 years). The commonest causes were gastro-enteritis (cholera) and septicaemia in 36.9% and 30.5% respectively. Twenty six (56.5%) of all the patients survived while the remaining 20 (43.5%) died. Twenty four (52.2%) patients had different complications of which pulmonary oedema was singularly found to significantly influence survival. Other factors that were found to significantly influence survival included availability of renal replacement therapy, the aetiology of ARF; gender; age of the patients and the duration of oliguria. We further compared the patients managed with haemodialysis with those managed with peritoneal dialysis and found no difference(s) in the age; duration of oliguria or hospitalisation; survival figures; effect of pulmonary oedema and the aetiology of ARF. However, the number of sessions for HD and the duration of PD significantly influenced survival. We conclude that ARF is still associated with a high mortality rate and prompt institution of available renal replacement therapy and aggressive management of complications would assist in reducing the trend.

The fetal intrahepatic umbilical vein as an alternative to cord needling for prenatal diagnosis and therapy.

Seventy-one fetal blood samplings (FBS) were attempted from the intrahepatic portion of the umbilical vein (IHV) at 18-34 weeks; 54 were attempted primarily and 17 secondarily after a failed attempt at the placental cord insertion. Fetal blood was obtained in 89 per cent of the cases. Intravascular transfusion (IVT) was attempted on 31 occasions and successful in 24 (77 per cent). In all cases of failed sampling or transfusion via the IHV, prenatal diagnosis and/or therapy was accomplished using alternative procedures. On only one occasion was the procedure postponed. There were no losses or neonatal morbidity attributable to the procedure. FBS from the IHV may be considered as an alternative approach to sampling the placental cord insertion. It is recommended in cases where the approach to the placental cord insertion is difficult or hazardous.