Seroprevalence and molecular detection of brucellosis among Pakistani women with spontaneous abortion.

Background: Human brucellosis is a neglected disease transmitted to humans from animals such as cattle, goats, dogs, and swine. The causative agents are bacteria of the genus Brucella, intracellular pathogens usually confined to the reproductive organs of their animal hosts causing sterility and abortions. The objective of the study was to determine the seroprevalence of brucellosis among women with spontaneous abortions (SAW) and compare this seroprevalence with that of healthy pregnant women (HPW). Methods: The case-control study was designed to determine the seroprevalence and molecular detection of brucellosis in women who suffered from spontaneous abortion and healthy pregnant women of the Haripur District of Pakistan. A total of 770 blood samples (n = 385 for each group) were collected from 9 public and 11 private hospitals in Haripur District from December 2021-March 2023. Data on demographic features, epidemiological variables, and risk factors were collected from each participant by structured questionnaires. Initial screening for brucellosis was performed by Rose Bengal Plate Test followed by qRT-PCR for molecular detection of the genus-specific BCSP-31 gene of Brucella. Results: The study showed that anti-Brucella antibodies were more found in SAW 23.63% (91/385) than in HPW 1.29% (5/385). Brucella specific DNA was amplified in 89.01% (81/91) seropositive samples of SAW. Demographic features and risk factors such as age, urbanicity, socioeconomic status, education, occupation, and animal contact were found significantly associated with brucellosis (p ≤ 0.05). Consumption of unpasteurized raw milk (OR = 18.28, 95%CI: 8.16-40.94) was found highly concomitant with seroprevalence. Conclusion: This study reports the first evidence of involvement of brucellosis in spontaneous abortions in women of Pakistan. The study can be used to develop strategies for risk management during pregnancy, to raise awareness for brucellosis, and develop control programs.

Pseudocitrobacter gen. nov., a novel genus of the Enterobacteriaceae with two new species Pseudocitrobacter faecalis sp. nov., and Pseudocitrobacter anthropi sp. nov, isolated from fecal samples from hospitalized patients in Pakistan.

Four isolates of Gram-negative facultatively anaerobic bacteria, three of them producing NDM-1 carbapenemase, were isolated from hospitalized patients and outpatients attending two military hospitals in Rawalpindi, Pakistan, and studied for their taxonomic position. Initially the strains were phenotypically identified as Citrobacter species. Comparative analysis of 16S rRNA gene sequences then showed that the four strains shared >97%, but in no case >98.3%, 16S rRNA gene sequence similarities to members of the genera Citrobacter, Kluyvera, Pantoea, Enterobacter and Raoultella, but always formed a separate cluster in respective phylogenetic trees. Based on multilocus sequence analysis (MLSA) including partial recN, rpoA, thdF and rpoB gene sequence and respective amino acid sequence analysis it turned out that the strains also here always formed separate clusters. Based on further comparative analyses including DNA-DNA hybridizations, genomic fingerprint analysis using rep- and RAPD-PCRs and physiological tests, it is proposed to classify these four strains into the novel genus Pseudocitrobacter gen. nov. with a new species Pseudocitrobacter faecalis sp. nov. with strain 25 CIT(T) (=CCM 8479(T)=LMG 27751(T)) and Pseudocitrobacter anthropi sp. nov. with strain C138(T) (=CCM 8478(T)=LMG 27750(T)), as the type strains, respectively.

Prevalence and molecular characterization of Enterobacteriaceae producing NDM-1 carbapenemase at a military hospital in Pakistan and evaluation of two chromogenic media.

The aim of this study was to assess the frequency and genotypic diversity of carbapenemase-producing Enterobacteriaceae (CPE) in stool samples from patients attending a military hospital in Pakistan. Further aims included the identification of factors that might predispose to faecal carriage and evaluation of 2 chromogenic culture media: Brilliance CRE and chromID CARBA. Of 175 patients, 32 (18.3%) had faecal carriage of CPE and all produced NDM-1 carbapenemase. All of these 32 patients were detected using chromID CARBA compared with 20 patients (62.5%) detected using Brilliance CRE (P = 0.0015). Duration of hospitalization and treatment with co-amoxyclav were statistically associated with a higher likelihood of carriage of CPE (P ≤ 0.05). The majority of NDM-1-producing Enterobacteriaceae co-produced CTX-M-1 group extended spectrum ß-lactamase, and one third produced armA-type methylase. NDM-1 carbapenemase was most commonly found amongst commensal types of Escherichia coli, especially phylogenetic group B1.

Ambu bag as a source of Acinetobacter baumannii outbreak in an intensive care unit.

This case report describes an outbreak of multidrug resistant Acinetobacter baumannii in the intensive care unit of a tertiary care hospital. Three patients were infected on the same day from an Ambu bag which was used on all the patients. The outbreak was immediately identified and the source was traced within one week. Appropriate measures were taken and a continuous surveillance was carried out resulting in reporting of no such case from the intensive care unit in the last 6 months.

Extended-spectrum ß-lactamase production in Shigella flexneri.

Emergence of multidrug-resistant strains of Shigella is a growing concern across the globe. Third-generation cephalosporins are used for treating infections caused by multidrug-resistant Shigellae. However, resistance to these cephalosporin antibiotics due to extended-spectrum ß-lactamases, has emerged as a new problem. So far extended-spectrum ß-lactamases producing Shigella has not been reported from Pakistan. We report such a case in Shigella flexneri from an 8-year old girl with acute dysentery.

Vancomycin resistant enterococcal sepsis in an acute myeloid leukemic patient.

Neutropenic fever is an important cause of morbidity and mortality during therapy of acute myeloid leukemia. Patients with acute myeloid leukemia are at increased risk of acquiring vancomycin resistant enterococcal infection and its treatment remains problematic. Vancomycin therapy for more than 7 days is usually associated with inducible vancomycin-resistant enterococcal infections. We report a case of vancomycin resistant enterococcal sepsis in a patient of acute myeloid leukemia.

Comparison of two different sensitivity testing agars for detecting methicillin resistance in Staphylococcus aureus.

OBJECTIVE: To compare the accuracy of Mueller-Hinton agar and Isosensitest agar using cefoxitin disc for detecting methicillin resistant Staphylococcus aureus using mecA gene PCR assay as gold standard. STUDY DESIGN: Comparative study. PLACE AND DURATION OF STUDY: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from May 2006 to January 2007. PATIENTS AND METHODS: One hundred clinical isolates of Staphylococcus aureus were evaluated; 64 MRSA (methicillin resistant Staphylococcus aureus) and 36 MSSA (methicillin sensitive Staphylococcus aureus) by mecA PCR assay. All the isolates were tested with cefoxitin 30 microg disc using semi-confluent growth on Mueller-Hinton agar as well as on Iso-sensitest agar in ambient air at 35-37 degrees C after an overnight incubation as per recommendations of Clinical and Laboratory Standard Institute. RESULTS: Following diameters provided the best sensitivity and specificity without substantial overlapping between the zones of resistant and sensitive isolates; Mueller-Hinton agar: R < or = 20 mm (sensitivity 100% and specificity 100%), S > or = 22 mm (sensitivity 97.2% and specificity 100%), and Iso-sensitest agar: R < or = 26 mm (sensitivity 100% and specificity 100%), S > or = 26 mm (sensitivity 100% and specificity 100%). High accuracy was obtained with cefoxitin disc on both media. CONCLUSION: Performance of both media was equally convincing for reliable prediction of methicillin resistance in Staphylococcus aureus by placing cefoxitin 30 microg disc on either of these in routine susceptibility testing.

Peripheral blood-based polymerase chain reaction in diagnosis of pulmonary tuberculosis.

BACKGROUND: The rapid diagnosis of infectious diseases, particularly those that represent a public health problem, like tuberculosis, is a challenging problem. By using nucleic acid amplification techniques like PCR, one may be able to diagnose, the disease on the day of arrival of specimen in the laboratory. For diagnosis of tuberculosis by direct methods like PCR, specimens from site of infection are required. In certain cases it is difficult to get the specimens from site of infection and in such situations; some researchers have tried to detect the DNA of Mycobacterium tuberculosis complex from blood of these patients. The purposive of this study is to determine the diagnostic efficacy of peripheral blood-based polymerase chain reaction for diagnosis of pulmonary tuberculosis. METHODS: This was a simple descriptive study, carried out in Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi from Jan 2004 to Dec 2004. Sputum and blood samples were collected from 96 suspected patients of pulmonary tuberculosis. Sputum samples processed for ZN staining and AFB culture (gold standard) and blood samples processed for PCR. RESULTS: Out of 96 cases, 60 (62.5%) were culture positive. PCR was positive in 14 (14.5%). AFB smear positive were 34 (35.4%). The overall sensitivity and specificity of the PCR assay was 20% and 94.4% respectively and the positive and negative predictive values were 85.71% and 41.46% respectively. The overall efficiency of the test was 47.91%. CONCLUSION: Due to low sensitivity; a negative PCR assay does not rule the disease. However, this test may be helpful in cases where specimens from the site of infection are not available.

Pneumocystis carinii and Trichosporon beigelii pneumonia following allogeneic haemopoeitic stem cell transplantation.

Pneumocystis Carinii and Trichosporon beigelii are opportunistic infections in immunocompromised patients. We report a case of a young lady who underwent haemopoeitic stem cell transplantation for relapsed acute lymphoblastic leukemia. This 25 years old female developed fever, dry cough and rapidly progressive dyspnoea during post transplant neutropenia and was found to be suffering from Pneumocystis carinii pneumonia. She was successfully treated with Co-trimoxazole. The patient again presented with similar symptoms on day 55 post transplant. This time Trichosporon beigelii was isolated from bronchoalveolar lavage and she responded to prompt antifungal therapy. Other complications encountered during the subsequent course were extensive subcutaneous emphysema and spontaneous pneumothorax that required chest intubation and brief hospitalization. The patient is presently nine months post transplant and is asymptomatic.