Latex Agglutination Research Articles

Nano magnetic-based ELISA and nano magnetic-based latex agglutination test for diagnosis of experimental trichinellosis.

Human trichinellosis is a worldwide foodborne public health threat. Detecting circulating antigens of Trichinella spiralis "T. spiralis" allows for an early diagnosis before larval encystation develops in skeletal muscles. For the first time, the present study aimed to formulate an effective nanomagnetic beads based-ELISA and -latex agglutination test (NMB-ELISA and NMB-LAT) to recognize T. spiralis adult worm crude extract antigen (AWCEA) in sera of experimentally infected mice. The study included thirty-eight mice classified into 3 groups; T. spiralis-infected group (GI) which was euthanized 6, 8, 10, 12, 14days post-infection (dpi), other parasitic infections group (GII) and healthy control group (GIII). Rabbit anti-T. spiralis polyclonal antibodies (pAbs) were utilized to detect AWCEA in serum samples by sandwich ELISA, NMB-ELISA, and NMB-LAT. Using NMB-ELISA, AWCEA was detected in sera collected at 6 and 8 dpi, with a sensitivity of 50% and 75%, respectively, and a specificity of 100%. Whereas, sandwich ELISA and NMB-LAT couldn't detect the antigen at the same time intervals. Both ELISA formats were able to detect the antigen in samples collected at 10, 12, and 14 dpi with a sensitivity of 100% for NMB-ELISA and 25%, 75%, and 100% respectively, for sandwich-ELISA. Yet, NMB-LAT couldn't detect AWCEA until 12 dpi with a sensitivity of 50% and specificity of 75%. In conclusion, NMB-ELISA is a promising sensitive tool for early and specific diagnosis of acute trichinellosis. The use of NMB-LAT could be a helpful screening procedure in field surveys.

Glycemic control as the main determinant factor of serum VEGF levels in type 2 diabetes mellitus patients.

Introduction. Diabetes mellitus (DM) is a main endocrine disorder that may cause vascular complications as the disease progresses. Vascular endothelial growth factor (VEGF) has been linked to the development of micro and macrovascular diabetic complications. This study aimed to assess several factors including blood pressure, body mass index, lipid profile, kidney function, and glycemic control that may provide the rise of serum VEGF levels in type 2 DM subjects. Methods. This cross-sectional study was carried out among 65 type 2 DM subjects. Systole, diastole, mean arterial pressure (MAP), and body mass index (BMI) were measured. The levels of serum VEGF were measured by Enzyme-linked immunosorbent assay (ELISA), Hemoglobin A1c (HbA1c) levels were measured by latex agglutination inhibition test, while serum glucose, lipid profiles, urea, and creatinine levels were tested by enzymatic photometric method. Results. The levels of serum VEGF had a significant correlation with BMI (p = 0.001, r = 0.397), fasting plasma glucose (FPG) (p = 0.001, r = 0.418), HbA1c (p < 0.001, r = 0.600), systole (p = 0.001), r = 0.397), diastole (p = 0.021, r = 0.286), and MAP (p = 0.001, r = 0.001). Further multivariate linear regression analysis revealed that HbA1c logarithm (log) was the determinant factor of VEGF levels (p < 0.001, β = 0.631, Adjusted R2 = 38.9%) Conclusion. HbA1c is the main determinant factor of serum VEGF levels among type 2 DM patients.

Cervicovaginal and rectal colonization by group B Streptococci in Mexican women with full-term pregnancies.

A pregnant woman rectally or vaginally colonized by group B Streptococcus can infect her newborn. Prospective, cross-sectional, analytical 24-month study in pregnant women. Women in labor with ≥ 36 weeks of gestation were included. Pregnancy was classified as normal or high-risk. Main risk factors of the pregnant women were analyzed. Rectal and vaginal samples were obtained, placed in Todd-Hewitt broth and subsequently inoculated in 5% sheep blood agar. Identification was carried out by biochemical tests and latex agglutination. 3,347 pregnant women were included. Mean age was 25.6 ± 5.3 years, 95.5% received antenatal care; 2,213 (66%) had normal-risk pregnancies, and in 1,370 (41%), delivery was by cesarean section. Overall colonization was 4.3% (145/3,347), and it was higher in the 30-34 years age group (6.8%). Serotype I (58%) was the most common. The percentage of colonization in this population was low. A routine cervicovaginal and rectal culture program in pregnant women and the intrapartum antimicrobial prophylaxis program are controversial in our region.

Serodiagnosis of map infection by latex agglutination test in slaugtered buffaloes of Malwa region (M.P., India)

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of chronic enteritis which is commonly called as Johne’s disease (JD) in animals which is associated with several incurable, auto-immune diseases like Crohn’s disease in human beings. In the current study, 19 serum samples of buffaloes were collected irrespective of their age, sex and breed which were slaughtered at Cantonment board slaughterhouse, Mhow and Nagar Nigam, Indore. These animals were brought from different places of Malwa region of Madhya Pradesh. For detection of anti-MAP antibodies, the latex agglutination test was performed following the standard procedure (Cheong Koo et al., 2004). The present investigation recorded 52.63% MAP infection in slaughtered buffaloes.

Workflow for Identification of Burkholderia pseudomallei Clinical Isolates in Myanmar.

Burkholderia pseudomallei, the highly infectious and causative organism of melioidosis, was first identified in Myanmar in 1911. B. pseudomallei was identified in Myanmar because of its genetic relatedness to Burkholderia species. In this study, we identified two isolates of Burkholderia cenocepacia, two Acinetobacter baumannii complexes, and 18 clinical isolates of B. pseudomallei using Vitek 2. These isolates were first screened using a latex agglutination test, which showed positive results in 20 of the 22 isolates. All isolates were cultured on Ashdown՚s agar and further tested using molecular methods. Specific PCR for type III secretion system (TTSs) gene clusters indicated 19 B. pseudomallei isolates out of 22 isolates. Furthermore, 16S rRNA and recA gene sequencing were used as the gold standard methods and yielded the same results. RapID NF Plus detected 16 B. pseudomallei out of 22 isolates. Vitek 2 and RapID NF Plus should be considered key tools in the diagnosis of melioidosis and surveillance of B. pseudomallei in Myanmar; however, accurate identification must be confirmed by TTS1 PCR. This study evaluated the presumptive workflow for the investigation of B. pseudomallei infections using different methods and options, in line with the available equipment.

Seroepidemiological study of Toxoplasnw gondif antiboCyamong women in Tikrit city

One hundred women were included in this study which referred by the physicians clinics for the detection of antitoxoplasma antibodies. Venous blood was collected from them, and serum obtained for the performed of rapid latex agglutination test and quelitative detective of Toxoplasma IgM by using Mercaptoethanol test.A questionnaire form was filled for each women by direct interview. The data requested inciuded age, occupation, residency, level of education, owing animals, marital status, history of abortion and numbers of abortion.Forty six (46%) women were dispiayed the presence of antitoxoplasma antibodies. Fifteen (32%) of the positive cases were positive for IgM antibodies.Significant association between the rate of total Toxoplasma antibodies and some epidemiological factors were included.

Comparative analysis of diagnostic methods for the detection of Cryptococcus neoformans meningitis.

Cryptococcosis is a devastating opportunistic infection in immunocompromised individuals, primarily in people living with HIV/AIDS. This study evaluated a protocol for the early diagnosis of meningitis due to C. neoformans, utilizing established molecular techniques from serum and CSF samples. The 18S and 5.8S (rDNA-ITS) sequence-specific nested PCR assays were compared with direct India ink staining and the latex agglutination test for detection of C. neoformans in serum and cerebrospinal fluid (CSF) from 49 Brazilian suspected meningitis patients. Results were validated with samples obtained from 10 patients negative for cryptococcosis and HIV, and by analysis of standard C. neoformans strains. The 5.8S DNA-ITS PCR was more sensitive (89-100%) and specific (100%) than the 18S rDNA PCR and conventional tests (India ink staining and latex agglutination) for identification of C. neoformans. While the 18S PCR exhibited a sensitivity (72%) similar to that of the latex agglutination assay in serum samples, it was superior to the latex agglutination assay when testing CSF, with a sensitivity of 84%. However, the latex agglutination was superior to the 18SrDNA PCR in specificity in CSF (92%). The 5.8S DNA-ITS PCR yielded the highest levels of accuracy (96-100%) of any test for detection (serological and mycological) of C. neoformans in both serum and CSF. Use of the nested 5.8S PCR was superior to other techniques for the diagnosis of cryptococcosis. The possibility of using serum, a non-invasively collected material, in a targeted 5.8S PCR analysis to identify Cryptococcus spp. is recommended, especially in immunosuppressed patients. Our results indicate that nested 5.8S PCR can increase the diagnostic capability of cryptococcosis, and we suggest its use to monitor patients in the future.

Dietary Fermentation Product of Aspergillus Oryzae Prevents Increases in Gastrointestinal Permeability (‘Leaky Gut’) in Horses Undergoing Combined Transport and Exercise

Equine leaky gut syndrome is characterized by gastrointestinal hyperpermeability and may be associated with adverse health effects in horses. The purpose was to evaluate the effects of a prebiotic Aspergillus oryzae product (SUPP) on stress-induced gastrointestinal hyperpermeability. Eight horses received a diet containing SUPP (0.02 g/kg BW) or an unsupplemented diet (CO) (n = 4 per group) for 28 days. On Days 0 and 28, horses were intubated with an indigestible marker of gastrointestinal permeability (iohexol). Half the horses from each feeding group underwent 60 min of transport by trailer immediately followed by a moderate-intensity exercise bout of 30 min (EX), and the remaining horses stayed in stalls as controls (SED). Blood was sampled before iohexol, immediately after trailering, and at 0, 1, 2, 4, and 8 h post-exercise. At the end of the feeding period, horses were washed out for 28 days before being assigned to the opposite feeding group, and the study was replicated. Blood was analyzed for iohexol (HPLC), lipopolysaccharide (ELISA), and serum amyloid A (latex agglutination assay). Data were analyzed using three-way and two-way ANOVA. On Day 0, the combined challenge of trailer transport and exercise significantly increased plasma iohexol in both feeding groups; this increase was not seen in SED horses. On Day 28, EX increased plasma iohexol only in the CO feeding group; this increase was completely prevented by the provision of SUPP. It is concluded that combined transport and exercise induce gastrointestinal hyperpermeability. Dietary SUPP prevents this and therefore may be a useful prophylactic for pathologies associated with gastrointestinal hyperpermeability in horses.

Nosocomial outbreaks of aseptic meningitis: A public health concern

Nosocomial outbreaks of aseptic meningitis: A public health concern

Inflammatory Markers in Chronic Kidney Disease

Background: Despite recent advances in chronic kidney disease (CKD) and end-stage renal disease (ESRD) management, morbidity and mortality in this population remain exceptionally high. Persistent, low-grade inflammation has been recognized as an important component of CKD, playing a unique role in its pathophysiology and being accountable in part for cardiovascular and all-cause mortality, as well as contributing to the development of protein-energy wasting. Material and methods; This study was conducted on CKD patients attending OPD &amp; IPD of Civil Hospital tarn taran. The participants having age more than 18 years &amp; less than 60 years. We assessed 120 individuals out of 60 are normal healthy individuals comprising the control groups &amp; rest 60 is of CKD cases. Serum CRP (mg/dl) concentration was measured by Latex agglutination test &amp; ESR (mm/hour) was measured by Wintrobe’s method. Serum Creatinine (mg/dl), Urea (mg/dl) &amp; Uric acid (mg/dl) concentration was measured by Modified Jaffe’s method, Urease &amp; Uricase method respectively. Results: - In the present study, serum CRP &amp; ESR was increased in CKD patients. The mean serum CRP levels of CKD patients &amp; controls were 33.55 ± 22.8 &amp; 2.07 ± 6121 respectively (p&lt; 0.001), highly significant result was observed. Mean level of serum ESR (40.25 ± 14.93) of cases shows statistical significant differences as compared with the mean of serum ESR of controls (13.50 ± 3.421). Conclusion: - CRP &amp; ESR are the markers used to evaluate kidney disease, however, each of these has its own limitation. The use of these inflammatory biomarkers may better assess overall patients’ risk &amp; be able to stage patients more appropriately.

Comparison between liquid chromatography/tandem mass spectroscopy and a novel latex agglutination method for measurement of hepcidin-25 concentrations in dialysis patients with renal anemia: A multicenter study

Background and aimsHepcidin-25 is an iron regulatory factor that plays an important role in anemia in patients with chronic kidney disease. Although liquid chromatography/tandem mass spectroscopy (LC-MS/MS) is the gold standard for measuring hepcidin-25 concentrations, results cannot be obtained immediately at clinical sites. In contrast, the latex immunoassay (LIA) can be performed using general clinical laboratory equipment, and the results can be obtained quickly. The aim of this study was to evaluate hepcidin-25 concentrations obtained by LC-MS/MS and a novel LIA method and compare the two methods. Materials and methodsHepcidin-25 was measured by LIA and LC-MS/MS in 182 hemodialysis patients. A hepcidin-25-specific reagent and an automatic analyzer were used for LIA, and a commercially available system was used for LC-MS/MS. The Passing-Bablok regression analysis method was used. ResultsOn Passing-Bablok regression analysis, the slope was 1.000, with an intercept of 0.359. Very strong associations were obtained, and the measured values were almost identical. ConclusionThe hepcidin-25 concentrations measured using LIA and those measured by LC-MS/MS were significantly correlated. LIA can be performed using general clinical examination equipment and has a higher throughput than LC-MS/MS. Therefore, measurement of hepcidin-25 concentrations by LIA can be useful for routine laboratory testing.

Association of the COVID-19 pandemic with changes in objectively measured sedentary behaviour and adiposity

BackgroundSeveral studies have reported that the coronavirus disease (COVID-19) pandemic has increased sedentary behaviour and obesity; however, these analyses used self-reported data, and the association between sedentary behaviour and visceral fat and adipocytokines during the COVID-19 pandemic remains unclear. We aimed to investigate the association of the COVID-19 pandemic with objectively measured sedentary behaviour and these obesity-related factors.MethodsLongitudinal analysis was conducted on 257 Japanese participants who underwent health check-ups in 2018 before and in 2020 during the COVID-19 pandemic. For both time points, sedentary behaviour was measured using an accelerometer for at least 7 days, visceral fat area (VFA) was measured using abdominal bioelectrical impedance analysis, and blood adiponectin level was measured using latex agglutination turbidimetric immunoassay. Multiple linear regression was performed to determine the association between sedentary behaviour and these outcomes.ResultsCompared with data in 2018, sedentary behaviour and VFA were significantly increased (P < 0.001, P = 0.006) whereas adiponectin level was significantly decreased (P < 0.001) in 2020. Increased sedentary behaviour was significantly associated with an increase in VFA (β = 3.85, 95% CI 1.22–6.49, P = 0.004) and a decrease in adiponectin level (β = −0.04, 95% CI −0.06 to −0.01, P = 0.005). However, the association of sedentary behaviour with adiponectin level was not significant after considering the effects of VFA.ConclusionsThe COVID-19 pandemic was associated with objectively measured sedentary behaviour and obesity-related factors in Japanese adults. Additionally, an increase in sedentary behaviour was associated with an increase in VFA, whereas the association of sedentary behaviour with adiponectin was partly mediated by VFA. These results suggest that avoiding increasing sedentary time is important to prevent visceral adiposity thereby ameliorating adiponectin, especially during behavioural limitations such as the COVID-19 pandemic.

Use of a phage cocktail to reduce the numbers of seven Escherichia coli strains belonging to different STEC serogroups applied to fresh produce and seeds

AbstractThe aims of this research were to evaluate the effectiveness of a phage cocktail at reducing seven Shiga toxigenic Escherichia coli (STEC) serogroups on different food matrixes: mung bean sprouts (MBP), lettuce, and mung bean seeds (MBS) and to test the phage cocktail effectiveness to reduce E. coli O157 on Romaine and iceberg lettuce. To study the effect of the type of food matrix on the STEC phage cocktail effectiveness, a mixture of seven highly sensitive STEC strains designated as phage propagation strains (PPS) were used to adulterate Romaine lettuce, MBP, and MBS matrixes at a concentration of 105 logs CFU/g. A subsample of the treated MBS was germinated to assess STEC survival. Recovered STEC strains were confirmed using latex agglutination and PCR. To test the phage cocktail effectiveness to reduce E. coli O157:H7 on Romaine and iceberg lettuce, a mixture of four STEC strains (different than phage propagation strains, non‐PPS) at both low (103 CFU/g) and high (105 CFU/g) concentrations were used to spike the samples in scaled up trials for the purpose of potential commercialization. Phage treatments including a combination of STEC phage cocktail and chlorinated water treatment were then applied to lettuce in a simulated scaled‐up trial. STEC was assessed on the treated samples at different storage time and temperatures (0, 24, 48, and 72 hr at 2, 10 and 25°C). In the food matrix trial, the combination of STEC phage cocktail and chlorinated water‐reduced PPS (p &lt; 0.001) STEC on lettuce by 2.1 log10 CFU/g and on MBP by 2.2 log10 CFU/g. However, isolates from all 7 STEC serogroups remained viable after phage treatment in both lettuce and MBP; particularly those associated with serogroup O111, O121, O103, and O145, while only a few colonies of serogroup O26, O45, and O157 were detected. Lettuce adulterated with low levels of 4 non‐PPS E. coli O157:H7 (103 CFU/g) achieved a reduction of 2.6–3.2 logs. While a reduction 1.7–2.3 logs was achieved by the phage cocktail when lettuce was inoculated with 105 CFU/g. Overall phage performance was more effective at 2 and 10°C and improved over storage time up to 72 hr. However, for MBS, the phage cocktail was not able to kill any of the STEC populations as all of them recovered during germination.

Latex Agglutination as an Alternative to the Hemagglutination Reaction of Influenza Viruses

As an alternative to the classical method of erythrocyte hemagglutination, a latex agglutination assay based on the interaction of influenza viruses with the sialoglycoprotein fetuin immobilized on the surface of polystyrene microspheres has been developed. Twelve influenza A virus strains of different subtypes and two influenza B viruses of different lines were tested. Simultaneous titration of viruses using the classical hemagglutination test and the proposed latex agglutination assay showed similar sensitivity and a high degree of correlation (R = 0.94). The obtained microspheres can be used for titration of viruses that recognize and bind sialylated glycans as receptors. In particular, latex aggregation was also induced by the Newcastle disease virus.

Invasiveness potential of pneumococcal serotypes in children after introduction of PCV13 in Blantyre, Malawi

IntroductionThe introduction of PCV13 to the Malawi infant immunization schedule in 2011 has been associated with reduced disease from Streptococcus pneumoniae. Improved understanding of serotypes with high invasive potential can guide future vaccination interventions. We aimed to estimate pneumococcal serotypes associated with acute respiratory infection (ARI) and invasive pneumococcal disease (IPD) in hospitalized children in Blantyre, Malawi.MethodsWe analysed data from healthy children under 5 years in the community in Blantyre and children admitted to Queen Elizabeth Central Hospital with ARI between 2015 and 2018. Nasopharyngeal swabs from children were tested for S. pneumoniae and serotyped by latex agglutination if positive. We analysed culture-positive blood and cerebrospinal fluid samples from admitted children between 2012 and 2018 to identify cases of IPD after the introduction of PCV13. We calculated the age-adjusted odds ratio (OR) of carriage for S. pneumoniae vaccine serotypes (VT) comparing those with ARI to healthy children. We also calculated age-adjusted ORs comparing serotypes causing IPD to carriage in the community with OR > 1 indicating high invasive potential.ResultsSerotypes 5 (OR 24.73 [95% CI 7.90–78.56] p < 0.001), 1 (OR 23.38 [95% CI 9.75–56.06] p < 0.001), and 6B (OR 4.73 [95% CI 1.66–11.64] p = 0.001) had high invasive potential. Serotype 6B was no longer significant (OR 1.34 [95% CI 0.07–6.87] p = 0.777) in a sensitivity analysis accounting for year of recruitment. The prevalence of S. pneumoniae carriage in the community was 72.6% [95% CI 71.3–74.0] (3078/4238) and 23.4% (719/3078) of positive community samples were VT. The carriage prevalence in those hospitalised with ARI was 45.5% [95% CI 42.1–48.9] (389/855) and 43.8% of hospital attendees reported antibiotic use prior to admission. We did not identify significant associations with carriage of any serotypes in those with ARI.ConclusionsPneumococcal serotypes 5 and 1 are associated with high invasive potential. Despite high community pneumococcal carriage, pre-hospital antibiotic usage likely reduces pneumococcal detection among children admitted in this setting and further research is needed to investigate serotypes associated with ARI. Data from this study can guide future preventative vaccination strategies in Malawi.

Staphylococcus aureus in Horses in Nigeria: Occurrence, Antimicrobial, Methicillin and Heavy Metal Resistance and Virulence Potentials

Staphylococcus aureus was isolated from a total of 360 nasal and groin skin swabs from 180 systematic randomly-selected horses slaughtered for meat at Obollo-Afor, Enugu State, Southeast Nigeria and antimicrobial, methicillin and heavy metal resistance profile and virulence potentials of the isolates established. Baird-Parker agar with egg yolk tellurite was used for S. aureus isolation. S. aureus isolates were confirmed biochemically and serologically using a specific S. aureus Staphytect Plus™ latex agglutination test kit. The antimicrobial resistance profile, methicillin, vancomycin and inducible clindamycin resistance, and β-lactamase production of the isolates were determined with disc diffusion. Tolerance to Copper, Cadmium, Lead and Zinc was assessed using the agar dilution method and virulence potentials were determined using phenotypic methods. Forty-three (23.9%) of the 180 horses harbored S. aureus. Some 71 S. aureus were recovered from the 360 samples. Two (2.8%) of the 71 S. aureus were methicillin-resistant S. aureus (MRSA) and 69 (97.2%) were methicillin-susceptible. MRSA was recovered from 2 (1.1%) of the 180 horses. Some 9.4% of the isolates were multiple drug-resistant (MDR). The mean multiple antibiotic resistance indices (MARI) for the isolates was 0.24. Heavy metal resistance rate of the isolates ranged between 35.4-70.4%. The isolates, including the MRSA strains, displayed virulence potentials as clumping factor and catalase, gelatinase, caseinase, heamolysin, and biofilm was at the rate of 100%, 53.5%, 43.7%, 18.3% and 23.9%, respectively. This study showed that a considerable percentage of horses slaughtered in Obollo-Afor Southeastern Nigeria are potential reservoirs of virulent multiple drug- and heavy metal-resistant S. aureus, including MRSA, that could spread to humans and the environment.

SEROPREVALENCE OF Toxoplasma go¬ndii IgG ANTIBODY IN WOMEN SUFFERING FROM ABORTION AT SANA’A CAPITAL AND CAPITAL TRUSTEESHIP

Toxoplasma gondii, is obligate intracellular protozoan, infects humans and any other warm-blooded animal. Toxoplasmosis is probably one of the most common human infections and becomes a life threatening disease in case of congenital infection or immunodepressed patient. The aim of this study was to estimate toxoplasmosis prevalence among Sana’a capital and capital trusteeship women suffering from abortion and to emphasize the consequences. The study was performed between 2006 and 2007 including women suffering from abortion. Latex agglutination (LAT) was used for Toxoplasma IgG detection. The result indicated that of 508 women suffering from abortion, 344 (67.7 %) were seropositive for Toxoplasmosis. The age (21-30 years) and (31-40 years) had the highest percentage (68.4 %) and (68.5 %) respectively followed by age (≤ 20 years) (66.0 %) of positive results. The most prevalent titers of LAT were 1/16 (25.0 %) followed by 1/32 (19.10 %) then 1/8 (8.30 %). The results confirmed Toxoplasma gondii wide spread at Sana’a capital and capital trusteeship and the risk represented by this protozoan. Therefore any raiser for this disease should be taken into consideration in order to diminish the consequences of this infection.

Clinical case of isolation and identification of &lt;I&gt;Streptococcus equi subsp. Zooepidemicus&lt;/I&gt;

Streptococci manifest their pathogenic properties by reducing the overall resistance of the organism or individual tissues.Purpose of the work: isolation, identification and study of the biological properties of isolates isolated from the foal.A pure culture was obtained, morphological, cultural and biochemical properties were studied.For the subsequent identification of streptococci to species, two api 20 Strep test systems (BIOMERIEUX, France) were used. To determine the serological group of streptococci, a latex agglutination test was used using a kit for diagnosing streptococci of groups A, B, C, D, F, and G (OXOID, UK).Laboratory methods of diagnostics for the identification of causative agents of streptococcosis and their species differentiation have been studied. Virulent Streptococcus equi subsp. zooepidemicus caused abscesses in the area of the elbow joint of the foal. Isolation of bacterial cultures was carried out by the bacteriological method. Isolated Streptococcus equi subsp. zooepidemicus had the fermentation of a number of carbohydrates: sorbitol, there was no splitting of lactose, tregolose. Streptococcus equi subsp. zooepidemicus hydrolyzed esculin, no D-ribose fermentation was noted. However, the fermentation of carbohydrates is not a stable and clear sign, therefore it is not used for differentiation and identification of streptococci.When inoculated on blood agar with 5% sheep blood after cultivation for 24 hours at 37 0C, small gray-white colonies were found, surrounded by a wide transparent zone of beta-hemolysis. Catalase test is negative.As a result, it was found that the isolated culture belongs to the species Streptococcus equi subsp. zooepidemicus belonging to group C according to Lancefield.Modern laboratory diagnostics requires an integrated approach to identification, which includes bacteriological and serological methods - polyphase analysis. When identifying streptococci, the determination of their serological group according to Lancefield is of great importance. For this purpose, a latex agglutination reaction is convenient using a kit for diagnosing streptococci of groups A, B, C, D, F and G (OXOID, UK), coagglutination "Streptotest A, B, C, D, G" (AQUAPAST).

Comparative evaluation of some techniques used for the detection of rabies virus.

Neurotropic viruses in the family Rhabdoviridae, genus Lyssavirus, are what cause rabies, an acute, progressive, and highly lethal encephalomyelitis. Evaluation of the used diagnostic techniques to determine the most simple; rapid and accurate test for rabies virus (RABV) recognition in different specimens aiming to reach a rapid diagnosis as a step aid in the disease control and to prevent or even minimize the suspected hazard. The used techniques included an infection trial of Swiss mice with the mice-adapted challenge rabies virus followed by the detection of the virus in the infected mices' brains. Virus detection was carried out through the application BHK21 cell line infection; fluorescent antibody technique; latex agglutination test (LAT); direct enzyme-linked immunosorbent assay (ELISA); rabies antigen detection kit ELISA; conventional polymerase chain reaction (PCR). It was found that virus inoculation in mice and BHK21 cell lines needs 5-7 days with positivity of 90% and 100%, respectively. Rapid antigen kit was able to detect rabies antigen in mice brains suspension and BHK21 infected fluid within 3-5 minutes with percentages of 60% and 55.5%, respectively. In 1-1.5 hours, the direct fluorescent antibody method (DFAT) detected 90% and 100% of the rabies antigen in BHK21 cell line infection and brain impressions, respectively. Latex agglutination showed clear results with 88.8% with BHK21 infected fluid within 3-5 minutes while it did not carry out on brain emulsions to prevent falsely positive results brought on by the presence of tissue fragments. Conventional one-step PCR revealed 100% positivity with either brain or cell culture preparations within 2 days. Direct ELISA showed 88.8% positivity with BHK21 infected fluid with 1 day of work. Mice inoculation test, cell culture infection; DFAT and PCR are the most accurate techniques for the detection of RABV with a positivity of 90%-100% followed by LAT and ELISA with a positivity of 88.8%, and lastly, rabies antigen ELISA kit (RAK) with a positivity of 55.5%-60% taking in consideration the required time for each. In addition, the positivity % of the applied tests revealed their sensitivity and specificity.

Antimicrobial Profiles and Conventional PCR Assay of Shiga Toxigenic Escherichia coli O157:H7 (STEC) Isolated from Cattle Slaughtered at Bedele Municipal Abattoir, South West Ethiopia.

Shiga toxin producing Escherichia coli O157:H7 (STEC) is considered the most prevalent food borne pathogen that has gained increasing attention worldwide in recent years. A cross-sectional study was carried out at Bedele Municipal abattoir on cattle that were reported healthy from detailed ante-mortem inspections and having various body conditions scores. A total of 516 samples were collected and examined after enriched in modified peptone water. Following an enrichment, the samples were plated onto MacConkey agar and then onto Eosin methylene blue agar. Finally after a few similar procedures, 14 E. coli O157:H7 (STEC) isolates were confirmed through latex agglutination test. The collected data were analyzed using SPSS version 20 statistical software. This study finding revealed that the overall prevalence of E. coli O157:H7 out of 516 samples was found to be 2.7%. However, on sample type basis, the prevalence of E. coli O157:H7 from feacal samples, carcass swabs, butcher hand swabs and knife swabs were 4.7%, 3.3%, 1.1% and 1.1%, respectively. It was also found that that the prevalence of E. coli O157:H7 was significantly affected by age groups of slaughtered cattle (p<0.05). Moreover, in vitro antimicrobial susceptibility test result on average showed that almost all of E. coli O157:H7 isolates were highly susceptible to kanamycin and no resistance was shown to ciprofloxacin and gentamicin. Finally, the conventional PCR detection of stx1, st2 and hylA genes revealed that only 21.4% and 14.3% were found to contain stx1 and hylA genes respectively. To wrap up, this study showed that Shiga toxin producing E. coli O157:H7 (STEC) isolates were found with almost low overall prevalence rate from all sample sources in this study site. Therefore, improving abattoir facilities and slaughter house workers' personal hygiene are recommended to curtail E. coli O157:H7 meat contamination in this abattoir.