Pre-analytical stability of selected biochemical analytes in serum of horses and oxen stored at -20°C.

BACKGROUND: Delays between blood collection and analysis are inevitable, and samples are always stored in the refrigerator. The current study aimed to evaluate the stability of serum total cholesterol (TC), triglycerides (TG), total protein (TP), albumin and urea (URA) in horses and oxen after storage at -20°C. METHODS: Sera from apparently healthy 20 male horses and 20 oxen were obtained and aliquots of serum were divided into 3 portions. The first tube was used for baseline (T0) measurement of analyte values, whereas the other two tubes, T1 and T2, were stored at -20°C for 1 and 2 months, respectively, and analyte measurement was done. RESULTS: Results showed that the stability of TP (g/dL), URA (mg/dL) and TC (mg/dL) in oxen was statistically significant (p < 0.05). In horses, the stability of URA (mg/dL), TP (g/dL) and TG (mg/dL) were also statistically significant (p < 0.05). Additionally, URA and TC in oxen exceed TEa following measurement at T2 and TG in horses following measurement at T1 and T2. CONCLUSION: Laboratories should consider the storage temperature and time for specific analytes among animals. Therefore, stability studies at various storage temperatures and times are recommended to fully validate the stability of the analytes.

Stability of Some Biochemical Parameters in Sheep and Goat Serum Stored at -20℃.

Background: In Veterinary Medicine biochemical investigation of serum is widely used to aid diagnosis and treatment. However, delays usually happen between sampling and analysis. As a result, the serum is stored in refrigerators. In this regard, information on the effects of temperature and storage duration on the stability of the analyte is incomplete in general and its effect in sheep and goat serum is not described. Therefore, the objective of this study was to examine the stability of selected biochemical analytes from sheep and goat serum following storage at -20℃ for 2 months. Methods: Serum from 20 apparently healthy male 2-2.5 year-old sheep and goats was obtained and aliquots of serum from each sample were kept in three tubes. The first tube is for baseline (T0), which is done within an hour, while the other two (T1 and T2) are stored at -20℃ for 1 and 2 months, respectively. Total protein, albumin, urea, total cholesterol, and triglycerides were assayed. Results: The results revealed that storage temperature and duration for up to 2 months had no significant effect on any analytes except for urea in goats. The changes in terms of total observed error (TEo) for total protein; albumin and urea were greater than the acceptable values in both animals. Conclusion: Thus, further studies are required to assure alteration of analyte at various storage temperatures and duration. In addition, implementation of quality systems to achieve quality targets for analytes with greater TEo as compared to the established TEa is needed.

Occurrence of Aflatoxins in Poultry Feed in Selected Chicken Rearing Villages of Bishoftu Ethiopia.

Background: Aflatoxins (AFs) are major contaminants of feed used in the poultry industry that negatively affect animal and human health. In Ethiopia, previous studies on AFs mainly considered cattle feed and milk but scarce information exists for poultry feeds. Methods: The aim of this study was to determine the occurrence of AFs in poultry feed in selected chicken rearing villages of Bishoftu. The study was conducted from December 2018 to May 2019. Thirty-three compound poultry feed samples were collected and analyzed for aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and total AFs (AFT) using high performance liquid chromatography (HPLC). The moisture content of the samples was also determined. Results: The result indicated that 31 (94%) from a total of 33 samples were contaminated with AFs. The mean levels of AFB1, AFB2, AFG1, AFG2 and AFT were 70.11 µg/kg, 13.50 µg/kg, 88.55 µg/kg, 18.00 µg/kg and 190.18 µg/kg, respectively. This study found AFs at a level above the limit of FDA regulatory levels of 20 µg/kg in 25 (72.75%) samples for AFT and 22 (66.67%) samples for AFB1. The analysis of moisture content of the samples, ranges from 7.33% to 11.17%, indicating all were at optimal value (<12%). Conclusion: The study showed the high contamination of AFs in poultry feeds with optimal moisture content and hence further investigations are needed to address the cause. The study also supports the need for preventive strategies of AFs contamination in poultry feeds in Bishoftu.

Molecular Characterization of Eimeria Species in Broiler Chickens, Ethiopia.

Background: Eimeria is a parasitic organism causing coccidiosis, an enteric disease of major economic importance in poultry. The conventional methods for species identification of Eimeria have major limitations. Methods: Fresh fecal samples were randomly collected from 50 small and large-scale commercial broiler farms located in Adama, Bishoftu, Dukem, and Mojo towns. Polymerase chain reaction (PCR)-based assay was used for the differentiation of Eimeria species circulating among study sites and broiler farms. DNA was extracted from Eimeria oocytes using a DNeasy Tissue Kit. The extracted DNA templates and the genus-specific primers (Invitrogen) were used for the amplification of the ITS-1 region from seven Eimeria species of chicken. Descriptive statistical analysis and t-test were used to summarize and analyze the data. Results: The PCR result confirms that all the seven species of Eimeria were detected in both small and large-scale broiler farms. Prevalence variation was found among broiler farms and between study sites. The frequency of E. brunetti (P<0.006) and E. tenella (P<0.04) in the small-scale broiler farms was significantly higher compared to that of in large-scale farms. A significantly higher frequency of E. acervulina (P<0.03) and E. brunetti (P<0.03) was detected in broiler farms of Dukem and Mojo compared to the broiler farms in Bishoftu. The study also revealed that multiple infections of Eimeria species per sample are common in most farms. Among the evaluated small-scale broiler farms of Bishoftu, 80% showed up to 5 mixed species. In addition, about 33% of large-scale and 20% of small-scale broiler farms showed 6-7 mixed species. Conclusion: This study characterized all the seven Eimeria species and revealed that multiple infections of Eimeria species per sample are common in most of the evaluated farms. The current findings might be useful for future anticoccidial vaccine development and for effective chemoprophylactic and control strategies.

Applicability of commercial clinical chemistry test kits for horse serum.

OBJECTIVE: Validation of a test method is critical for confirming that the test can generate accurate and precise data. Although commercial biochemical test kits exist there are no specific and validated commercial clinical chemistry test kits designed for horses. The aim of this study was to validate commercial clinical chemistry test kits designed for a human serum for use in horses. RESULTS: Blood samples were collected from 29 apparently healthy adult male horses and pooled serum was prepared. Validation comprises replication and recovery experiments. Total observable error (TEo), sigma (σ) metrics, and quality goal index (QGI) were used to support the validation studies. Intra- and inter-assay variability was 2.05% and 2.08%, 2.26% and 1.89%, 2.4% and 1.63%, for total cholesterol, urea and total protein, respectively; recovery was 99.46%, 97.32%, and 100.1% for total cholesterol, urea and total protein, respectively. TEo% for the specified analytes was within the total allowable error (TEa). All three analytes satisfied the recommended requirement (> 3σ). The QGI for urea, as it had below 6σ was 0.95 indicating imprecision and inaccuracy. The results endorse the suitability of the studied commercial test kits and illustrated the acceptance criteria for horse's serum.

Xpert MTB/RIF assay for diagnosis of pulmonary tuberculosis in sputum specimens in remote health care facility.

BACKGROUND: Xpert MTB/RIF assay is considered as a great advance over conventional smear and culture in the diagnosis of TB and MDR-TB by simultaneously detecting M.tuberculosis and rifampicin resistance bacilli. However, very little information regarding the performance characteristics of Xpert MTB/RIF assay is available in Ethiopia. Therefore, the purpose of this study was to evaluate the performance of Xpert MTB/RIF assay compared to conventional sputum smear and culture methods for the diagnosis of pulmonary tuberculosis in remote health care facility. METHODS: A paired expectorated sputum samples were obtained from 227 consecutively recruited patients with signs and symptoms suggestive of tuberculosis at Karamara hospital during December 2013 to May 2014. One of the sputum specimen was tested directly by Ziehl-Neelsen staining and Xpert MTB/RIF assay without NALC-NaOH decontamination. The other of pair of sputa specimen was cultured for isolation of TB bacilli by conventional methods. Diagnostic performance of Xpert MTB/RIF assay and AFB smear microscopy were calculated against culture as the gold standard. RESULTS: Overall 25.5% (58/227) samples were positive for Mycobacterium tuberculosis complex (MTBC) by MGIT and/or LJ media of which 36.2% (21/58) and 65.5% (35/58) were positive by AFB smear microscopy and Xpert MTB/RIF respectively. The sensitivity, specificity, as well as the positive and negative predictive value of Xpert MTB/RIF assay were 65.5% (95% CI: 53.3-77.7%), 96.3% (95% CI: 93.4-99.2%), 86.4% (95% CI: 76.2-96.5%), and 88.6% (95% CI: 83.9-93.3%) respectively. Eighteen of 58 (31%) cases that were smear microscopy negative, were positive by Xpert MTB/RIF assay. CONCLUSIONS: Although Xpert MTB/RIF assay demonstrated high sensitivity in detecting MTBC in sputum specimens compared with conventional AFB smear microscopy, it demonstrated suboptimal sensitivity in smear negative patients compared to conventional culture.

Burden of metabolic syndrome among HIV-infected patients in Southern Ethiopia.

BACKGROUND: HIV infection and highly active antiretroviral therapy (HAART) can induce metabolic disturbances including lipodystrophy, dyslipidemia, and insulin resistance, which are reminiscences of metabolic syndrome (MS). However, little is known regarding the magnitude of MS in Ethiopian HIV population. This study, aimed to estimate the prevalence of MS among HIV positive patients with and without HAART. METHODS: A cross-sectional study was conducted at Hawassa University Referral Hospital, southern Ethiopia between February 2012 and April 2013. Data on demographic and anthropometric characteristics were collected from a total of 374 HIV positive participants (188 on ART and 186 on Pre-ART) using WHO stepwise approach. Fasting blood glucose, total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol was measured. The International Diabetes Federation (IDF) and the National Cholesterol Education Program: Adult Treatment Panel III (ATP) Criteria were used to define MS. RESULT: Of the 374 study participants 68% were females, and 50.3% were receiving ART. Using the IDF criteria, metabolic syndrome was diagnosed in 25% of patients receiving ART compared to 22.5% of the ART naïve group (OR: 1.14 CI: 0.71-1.84). Using the ATP criteria, the prevalence of MS was 18.1% in the ART groups compared to 15.6% in ART naïve group (OR: 1.20, CI: 0.69-2.06). Patients receiving ART had significantly elevated Cholesterol, triglyceride, glucose and LDL-c levels but lower CD4(+) cell counts than the Pre-ART groups. Being a female, having BMI of at least 25, older age (i.e. age≥45 years) and having total cholesterol of at least 200mg/dl were significantly associated with the presence of MS. Using the ATP criteria to define MS, taking d4T-3TC-EFV regimen was significantly associated with higher odds of MS. CONCLUSION: Almost a quarter of HIV patients on ART developed metabolic syndrome. Furthermore patients on ART had elevated lipid profile and glucose metabolism disturbance than the ART naïve.