Isolation and Molecular Characterization of <i>Streptococcus</i> <i>iniae</i> from African Catfish (<i>Clarias</i> <i>gariepinus</i>) from Selected Markets in Federal Capital Territory, Abuja, Nigeria

The Directigen 1-2-3 Group A Strep Test (DGAST; BBL Microbiology Systems, Cockeysville, Md.) was compared with conventional culture procedures on Trypticase soy agar with 5% sheep blood (BBL) and Selective Streptococcal Agar (ssA; BBL) for detection of group A beta-hemolytic streptococci (GABHS) for 1,006 patients complaining of sore throat. The DGAST was performed at five acute-care clinics according to the instructions of the manufacturer; interpretation of the cultures was done at the central microbiology laboratory. Of 924 patients with complete data, 243 (26.3%) were positive for GABHS on culture when both sheep blood agar and ssA were used. Of the patients with positive cultures, 159 were detected by the DGAST, yielding a sensitivity of 65.4%, a specificity of 84.7%, a positive predictive value of 60.5%, and a negative predictive value of 87.3%. The greater the number of colonies on culture, the greater the sensitivity of the DGAST, and the more intense the positive reaction on the DGAST, the higher the positive predictive value of the test. For the identification of GABHS, sheep blood agar was superior to ssA by 12.9% at 24 h and by 3.4% at 48 h of incubation.

The occurrence of Acetobacter diazotrophicus was directly demonstrated in plant tissues using a species-specific oligonucleotide probe and polymerase chain reaction (PCR) amplification of a 411-bp product. The oligonucleotide probe was derived from the sequence of a highly variable region of 23S rDNA and its specificity was tested with membrane-bound nucleic acids of 112 different microorganisms in hybridization experiments. It was found to be able to discriminate Acetobacter diazotrophicus from other Acetobacter spp. and other reference organisms. PCR amplification from pure cultured cells or colonies showed that the method was sensitive enough to detect as few as 200 cells in the reaction. The presence of Acetobacter diazotrophicus in tissues of micropropagated sugarcane plants inoculated with either this bacterium or a mixture of this bacterium and Herbaspirillum seropedicae was demonstrated by PCR amplification. Acetobacter diazotrophicus could also be detected by the PCR method in field-grown sugarcane plants, as well as in certain cultivars of Pennisetum purpureum Schumach but not in\i maize, sweet potato, and two samples of weed plants grown within or outside of a sugarcane field. The addition of 1% polyvinylpolypyrrolidone during preparation of the field samples, especially with root tissues, improved the amplificability of the target sequence. The minimum level of detection of this bacterium in sugarcane tissue using the universal 1440 and AD species-specific primers was about 105bacterial cells/g of fresh plant material. The sensitivity could be improved 10-fold by probing immobilized PCR products containing the target region with the32P-labeled oligonucleotide AD.Key words: Acetobacter diazotrophicus, diazotrophic endophytes, specific rRNA-targeting oligonucleotides, polymerase chain reaction (PCR).

Enhanced detection of methicillin-resistant Staphylococcus aureus using chromogenic media compared to traditional culture methods

Study of the Prevalence of Staphylococcus Aureus in Marine and Farmed Shrimps in Iran Aiming the Future Development of a Prophylactic Vaccine

Staphylococcus aureus is the most important pathogen of Sea foods. food poisoning happens in human due to consuming aqua products contamination with this bacteria and its enterotoxin. The procedure for maintenance and preserve the quality of these products are important for the production and growth of pathogenic bacteria and toxin production from fishing time and transporting them to stores until they are consumed. A total of 600 samples, containing 300 shrimps and 300 fishes( fresh and frozen ,farm and marine ) were studied, in order to detection of Staphylococcusaureus according to Iran National Standard From September 2013 to September 2014. Baird Parker agar containing egg-yolk and tellurite emulsion were used for isolation. Isolates were identified using the following criteria: production of coagulase, DNase, catalase, mannitol fermentation , hemolytic zone on 5% sheep blood agar, VP test and Gram staining. In total 206(34.3%) sample inclusive 84 shrimps (28%)and 122 fishes (40.7%) were contaminated with Staphylococcus aureus . Due to the presence of Staphylococcus aureus in fishes and shrimps, enforcement of quality control standards by the fisheries and careful monitoring of fishing and farming and preparation, freezing and transporting of marine products as well as health of workers are essential.

Buffalo (Bubalus bubalis) meat is a major item of export from India but export of beef i.e. meat from cattle (Bos indicus) is prohibited. Also, adulteration of buffalo meat with that of beef (meat from cattle) is a common fraudulent practice because of prohibition on cow slaughter in most states of India. Food analysts require precise identification techniques to implement such regulations. In the present study, a method of DNA extraction by Alkaline lysis from meat samples and speciation of buffalo meat using species specific Polymerase Chain Reaction (PCR) has been reported. Alkaline lysis technique is a rapid method which involves triturating meat with four volumes of 0.2N NaOH, dilution of resultant liquid extract with eight volumes of 0.2N NaOH, heating the mix 75°C for 20min followed by neutralization with eight volumes of 0.04N Tris HCl. Entire procedure of DNA extraction takes less than 30min and it is economical as it involves less expensive chemicals. Method was successfully applied in animal byproducts also viz., liver, heart and kidney. For authentication of buffalo meat, pair of primers was designed based on mitochondrial D loop gene nucleotide sequence. PCR amplification using the designed primers gave amplicon of size 482bp in buffalo and no amplification was detected in closely related species viz., cattle, sheep and goat meat samples. Results of the assay were highly repetitive and reliable. An export sample referred by export regulation authorities was also analyzed by using the Alkaline lysis method of DNA extraction and species specific PCR which enabled authentication of meat within 5h.

Stimulated by Hadley's 1 article on microbic dissociation, 12 tried by environmental conditions to incite a dissociation of measles cocci. No change in morphology or type of colony was demonstrated, but under the influence of normal and immune horse serum, and high temperature 40-41 C. for 24 hours, these cocci completely lost their specificity as determined by the opsonic method. Their specificity was quickly restored by transplanting them again onto blood agar and growing 'at 36 C. Recently a measles diplococcus was grown for 4 days instead of 1 day at 40 C. and although it was transferred for 22 generations on blood agar at 36 C, its specificity was not restored. Again no change in morphology or type of colony was observed. After some strains of measles diplococci have been isolated several years the growth on blood agar is seen to be less green, moister and more abundant, with a partial zone of hemolysis around the colonies, while the cocci grow in clumps and pairs and now show no chain formation. The cocci are more phagocytable, so that it is necessary to use very thin suspensions in order to demonstrate differences between the opsonic specimens containing normal and immune serum. One strain which for years grew in this way, when cultured for 24 hours in oxalated sheep blood and dextrose broth equal parts, repeatedly developed long chains and capsules. In this medium many of the cocci were seen inside leukocytes. When plated on sheep blood agar, two types of colonies were seen, a fairly large smooth, green colony with a narrow zone of partial hemolysis and a large, smooth, moist, less green colony without the zone of hemolysis. The cocci from the large hemolytic colony were more in clumps, those in the large nonhemolytic colony were more in pairs; the cocci from the former colony were much more phagocytable than those from the non-hemolytic colony, in the presence of both normal and immune serum. This change or dissociation appeared to be due to the presence of fibrin with leukocytes in

The study was carried out to determine the phenotypic characters of Aeromonas isolates from fresh water fishes sold in FCT using culture and isolation, and conventional biochemical identification method and confirmation with MicrobactTM GNB 24E identification system. Out of 400 samples collected from different fishes (257 from Tilapia zillii, 77 from Clarias gariepinus, 58 from Lates niloticus, and 8 from Alestes nurse) culture and biochemical characterization revealed that 40 isolates were Aeromonas specie and all the isolates (100%) were positive to oxidase, catalase, hydrogen sulphide, voges proskauer and motility tests. MicrobactTM GNB 24E kit further revealed 15 out of 40 isolates were Aeromonas hydrophila. Descriptive statistics of the results showed an overall prevalence rate of 3.75% with the highest prevalence rate of 6.79% in Bwari Area Council, 3.29% in Abuja Municipal Area Council. Aeromonas hydrophila was isolated from all the species of fish in the study area. Tilapia zillii had a prevalence rate of 3.11%, while Lates niloticus 5.17%, Clarias gariepinus, 3.89%, and Alestes nurse was 12.50%. Our research was able to isolate Aeromonas hydrophila from fresh water fish sold in Federal Capital Territory, which causes zoonotic diseases therefore implies potential danger to man and other animals which makes public health awareness and enlightenment of the dangers associated with Aeromonas hydrophila in Nigeria necessary.

Fish species are important aquatic models utilised in ecotoxicology studies; however, most of these species are found in temperate countries. In this chapter, we reviewed native fish species in Nigeria that are utilised for ecotoxicology studies. A search of the literature was conducted using Google Scholar search engine from anytime until November 7, 2023, resulting in a total of 45 articles that were included in this review. The commonly used native Nigerian fish species in laboratory and field-based (biomonitoring) ecotoxicological studies are the African sharptooth catfish Clarias gariepinus, the Nile tilapia Oreochromis niloticus and the Guinean tilapia Coptodon guineensis. These fish species have been demonstrated to be valid models to study biomarkers of exposure to and effects of toxicants including pesticides, effluents, heavy metals, hydrocarbons, pharmaceuticals, plant and sediment extracts. They serve as bioindicators of stressors or pollutants in biomonitoring programmes with indices ranging from cellular (nuclear abnormalities, haematological effects) to tissue and organ histological alterations, oxidative stress indices as well as individual/population level effects (such as behavioural changes). We recommend targeted studies on the culture exploitation of other native Nigerian freshwater, brackish and marine fish species as well as ratification of C. gariepinus and O. niloticus as globally recognised model fish species for use in ecotoxicological studies.

Background: Fish is known to be one of the cheapest sources of animal protein and have essential nutrients needed in human diets. The present study investigated the proximate and nutrient composition of three species of fish, Champsocephalus gunnari, Oreochromis niloticus, and hybrid catfish (Clarias gariepinus). Methods: The sample collection, proximate and mineral analysis were conducted using the standard protocols of sample collections and chemical analysis. Results: The results revealed that Champsocephalus gunnari, Oreochromis niloticus, and Clarias gariepinus contain moisture (66.00±0.50%, 52.00±1.00%, and 70.00±0.20%), crude protein (9.20±0.1323%, 3.75±0.02%and 5.80±0.05%), crude lipid (10.16±0.91%, 2.37±0.01% and 12.00±0.30%), ash (11.92 ±0.02%, 39.40±0.03% and 11.97±0.06%), and crude fiber (2.03±0.01%, 2.36±0.02% and 0.19±0.01%) respectively. The mineral contents were: iron (4.50±0.01mg/kg, 3.70±0.01 mg/kg and 4.70±0.02mg/kg), Zinc (2.35±0.01mg/kg, 2.15±0.0100mg/kg and (1.89±0.01mg/kg) for Champsocephalus gunnari, Oreochromis niloticus, and Clarias gariepinus respectively, while copper was only detected in C. gunnari (0.25±0.01mg/kg). The same amount of chromium (0.01±0.00 mg/kg) was detected in all the samples. Manganese was undetected in all the samples. Conclusion: In conclusion, the present study demonstrated the nutritional value of Champsocephalus gunnari, Oreochromis niloticus and Clarias gariepinus. This information would help in choosing any of the fish bases on their nutritional values rather than taste and other physical features.

'Query' (Q) fever is a neglected but emerging or re-emerging zoonotic disease caused by the bacterium Coxiella (C.) burnetii. Several host species are considered or speculated to be the primary reservoir hosts for human infection. In the past, several research groups in Nigeria have evaluated the prevalence of C. burnetii in various vertebrate and invertebrate hosts. Currently, there is a paucity of knowledge regarding the epidemiology of the pathogen in Nigeria with limited or no attention to control and prevention programs. Therefore, this review was undertaken to comprehend the current situation of C. burnetii infection in human, domestic and peri-domestic animals, and some tick species in Nigeria since 1960 with the aim to help identify future research priorities for the country. A comprehensive literature search was performed using the PRISMA guidelines on five scientific databases including Google Scholar, PubMed, AJOL, Science Direct, and Scopus for articles published from Nigeria dealing with the screening of blood, milk, or tick DNA for evidence of C. burnetii using any standard diagnostic approach. Of the 33 published articles subjected to full-text evaluation, more than 48% of the articles met the inclusion criteria and were thus included in this review. We observed different ranges of prevalence for C. burnetii antibodies from four vertebrate hosts including cattle (2.5-23.5%), sheep (3.8-12.0%), goats (3.1-10.9%), and humans (12.0-61.3%). Additionally, the use of molecular diagnostics revealed that the DNA of C. burnetii has been amplified in eight tick species including Hyalomma (Hy) dromedarii, Hy. truncatum, Hy. impeltatum, Hy. rufipes, Hy. impressum, Amblyomma (Am.) variegatum, Rhipicephalus (Rh.) evertsi evertsi, and Rh. annulatus. Two rodent's species (Rattus rattus and Rattus norvegicus) in Nigeria were documented to show evidence of the bacterium with the detection of the DNA of C. burnetii in these two mammals. In conclusion, this review has provided more insight on the prevalence of C. burnetii and its associated host/vector in Nigeria. Domestic animals, peri-domestic animals, and ticks species harbor C. burnetii and could be a source of human infections. Due to the paucity of studies from southern Nigeria, we recommend that research groups with interest on vector-borne diseases need to consider more epidemiological studies in the future on C. burnetii prevalence in diverse hosts to help unravel their distribution and vector potentials in Nigeria as a whole.

Clarias gariepinus is an important commercial fish species in Nigeria and is widely cultivated by many farmers. This study investigated the effects of dietary supplementation with Chlorella vulgaris on liver biomarkers and lipid profiles in Clarias gariepinus over 12 weeks. Five groups of fish were used in the study; one group served as the control, while the other four groups were fed different concentrations of the algae at 5%, 10%, 15%, and 20%. At the end of the feeding trial, the fish were anesthetized using clove oil, and blood samples were collected for analysis. The results indicated that the supplementation of C. vulgaris in the diets of C. gariepinus led to higher protein percentages (ranging from 43.0% to 47.3%) and lower levels of AST (230.0) and ALT (54.0) in the algae-fed groups compared to the control group, which had a protein percentage of 39.67, AST levels of 497.3, and ALT levels of 180.7. Additionally, the lipid profiles showed that the treated groups had better outcomes than the control group. The findings revealed significant variations in the fish's serum biochemical parameters and lipid profiles across the different treatments. Therefore, this study highlights the positive impact of microalgae as a physiological booster in C. gariepinus.

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Introduction: Milk handling in the Nigerian informal dairy sector is commonly done without observing hygienic practices, which is a threat to food safety and public health. The indiscriminate use of antibiotics in treatment of infections in animals has led to increasing resistance of pathogenic organisms to commonly used antibiotics. We determined the bacteriological quality of nono and the antimicrobial resistance of respective bacterial isolates. Methods: We conducted a cross-sectional study on 300 samples of ready-to-consume locally fermented milk product (nono) sold in the Federal Capital Territory, Nigeria between June and September 2018. We used a multistage sampling technique for sample collection. Culturing on appropriate media and conventional biochemical tests were carried out for identification and isolation of bacteria of interest while further confirmatory tests were carried out using Microbact/Mocrogen® kits. Serological test was conducted using Remel RIM™ Latex to confirm presence of E. coli 0157:H7 sero-group, while Kirby-Bauer disc diffusion method was used for antimicrobial susceptibility testing. Results: Total Aerobic Plate Count (±SD) ranged from 5.6 ± 1.7 - 7.0 ± 0.4 log10 cfu/ml while total coliform count ranged from 5.6 ± 0.5 - 6.5 ± 0.7 log10 cfu/ml. Out of the 300 samples, 37 (12.3%) tested positive for E. coli out of which 16% were of the 0157:H7 sero-group while 21(7%) were positive for Staphylococcus aureus. E. coli isolates were found to be totally resistant to Vancomycin and Methicillin, and almost totally resistant to Ampicillin (97%) and Tetracycline (95%). Staphylococcus aureus isolates on the other hand were totally resistant to Oxacillin, Ticarcillin and Amoxycillin. Conclusion: Total plate counts from this study is above the maximum permissible range of 4.6 log10 cfu/ml. The bacterial isolates showed multidrug resistance varying from 3 to 8 of the antibiotics used. We recommended health education and awareness creation on hygiene practices among local milk vendors and advocated for right use of antimicrobials in animals by veterinarians.

The fatty acid profile of feral and cultured Heterobranchus longifilis, Clarias gariepinus and feral Chrystichthys nigrodigitatus were determined. Three adult fish each of Heterobranchus longifilis and Clarias gariepinus were procured from Oguta lake in Imo State and Azuberth Research Complex at Industrial Cluster Naze, in Owerri west Loca Government Area, Imo State, while Chrystichthys nigrodigitatus were obtained from Afikpo Ebonyi State. The fish samples were analyzed in the laboratory to determine the, fatty acid profile using recommended standard methods by AOAC. Data obtained were then subjected to statistical analysis and means separated for comparison. Thirteen fatty acids were obtained in all the treatments (H. longifilis, C. gariepinus and C. nigrodigitatus both culture and wild respectively. Myristic, lauric, stearic and palmitic acids were the saturated fatty acid found. While Linolenic acid, Tetracopentaenoic, Linoleic acid, Arachidonic acid, Cervonic acid, Oleic acid, Dihomo-linoleic, Linolenic acid and Eicosapentaenoic acid are the unsaturated fatty acid. However, Linoleic acids (C18:2) had the highest concentration (28.160%) in wild H. longifilis, followed by cultured C. gariepinus (15.668%). Generally, when pooled together for each fish species unsaturated fatty acid had far higher percentage concentration than saturated fatty acids. The presence of linoleic acid (Omega fatty acid) in high amount in all the treatments is an indication that fish oil is an excellent source of polyunsaturated fatty acid that the body cannot synthesized and must be required in their diet hence, making fish a must for human consumption. Though saturated fatty acids were present but the concentrations of beneficial unsaturated fatty acids far outweighs the presence of the low concentration of saturated fatty acids.