A three year retrospective study on the increasing trend in seroprevalence of dengue infection from southern Odisha, India.

BACKGROUND & OBJECTIVES: In Odisha, several cases of dengue virus infection were detected for the first time in 2010, the importance of dengue as a serious mosquito-borne viral infection was felt only in 2011 with the reporting of many more positive cases. This retrospective three year study was done to find out the seroprevalence of dengue Ig m0 antibody and to know the predominant serotype of dengue virus among the patients suspected to have dengue virus infection in a tertiary care hospital in southern Odisha, India. METHODS: Blood samples from clinically suspected dengue cases admitted in the Medicine and Paediatrics departments of a tertiary care hospital were collected. These were processed for detection of dengue specific IgM antibody, carried out by the ELISA method. Dengue IgM antibody positive serum samples were tested for serotypic identification. RESULTS: Of the 5102 samples tested, 1074 (21.05%) were positive for dengue IgM. Maximum numbers of cases were found in 2012. Majority (47.86%) of cases were detected in the month of September. The most common affected age group was 11 to 20 yr. DENV1 and DENV2 were the detected serotypes. INTERPRETATION & CONCLUSIONS: Rapid increase in the dengue cases in 2012 became a public health concern as majority of cases were affecting the young adolescents. Most of the cases were reported in post-monsoon period indicating a need for acceleration of vector control programmes prior to arrival of monsoon.

Sequence analysis of pfcrt and pfmdr1 genes and its association with chloroquine resistance in Southeast Indian Plasmodium falciparum isolates.

BACKGROUND: Due to the widespread resistance of Plasmodium falciparum to chloroquine drug, artemisinin-based combination therapy (ACT) has been recommended as the first-line treatment. This study aims to evaluate the extent of chloroquine resistance in P. falciparum infection after the introduction of ACT. This study was carried out based on the mutation analysis in P. falciparum chloroquine resistant transporter (pfcrt) and P. falciparum multidrug resistance 1 (pfmdr1) genes. Identification of these molecular markers plays a significant role in monitoring and assessment of drug resistance as well as in designing an effective antimalarial drug policy in India. METHODS: Sixty blood samples were collected from patients infected with P. falciparum from JIPMER, Puducherry and MKCG Medical College, Odisha. Polymerase chain reaction-restriction fragment length polymorphism was performed, targeting the point mutation of K76T in pfcrt and N86Y in pfmdr1 gene. The PCR products were sequenced, genotyped and further analysed for amino acid changes in these codons. RESULTS: The frequency of pfcrt mutation at 76th position was dominant for mutant T allele with 56.7% and wild type K, 43.3%. Majority of pfmdr1 86 allele were wild type, with N (90%) and mutant, Y (10%). Additionally, we found three haplotypes for CQ resistance, SVMNT, CVIET and CVIKT in association with the pfcrt gene. However, a poorly studied SNP in pfmdr1 gene (Y184F) associated with CQ resistance showed high frequency (70%) in P. falciparum isolates. CONCLUSIONS: The point mutation K76T of pfcrt is high in P. falciparum suggesting a sustained high CQ resistance even after five years of the introduction of ACTs for antimalarial therapy. The present study suggests a strong association of CQ resistance with pfcrt T76, but not with the pfmdr1 Y86 mutation. However, sequence analysis showed that Y184F mutation on pfmdr1 gene was found to be associated with high resistance. Also, a new pfcrt haplotype 'CVIKT' associated with CQ resistance was found to be present in Indian strains of P. falciparum. The data obtained from this study helps in continuous monitoring of drug resistance in malaria and also suggests the need for careful usage of CQ in Plasmodium vivax malarial treatment.

Primary actinomycosis of hand.

Actinomycosis is a chronic granulomatous suppurative disease having the propensity for extension to the contagious tissue with the formation of multiple discharging sinus tracts. Primary actinomycosis of extremity is a very uncommon clinical entity and is commonly considered as a soft-tissue infection. We report here, a case of primary actinomycosis of the upper extremity in a 24-year-old male who was treated successfully with surgical excision and extended period of antimicrobial treatment.

Prevalence of cryptococcal meningitis among people living with human immunodeficiency virus/acquired immunodeficiency syndrome in a Tertiary Care Hospital, Southern Odisha, India.

OBJECTIVE: Cryptococcal meningitis (CM) caused by encapsulated opportunistic yeast Cryptococcus neoformans is an important contributor to morbidity and mortality in people living with human immunodeficiency virus/acquired immunodeficiency syndrome (PLHAs). Early diagnosis of such patients is the key to their therapeutic success. A retrospective study was conducted to evaluate the clinical features, laboratory findings, and prevalence of CM among hospitalized PLHAs in a tertiary care setting. MATERIALS AND METHODS: A total of 112 clinically diagnosed CM patients were subjected to cerebrospinal fluid analysis and tests for human immunodeficiency virus antibodies by the standard laboratory operating procedures. RESULTS: Out of 112, 16 showed a definite diagnosis of C. neoformans with the prevalence of 14.3%. Males in the age group of 21-40 years were most commonly affected than females. The clinical manifestations observed were fever and headache (100%), followed by altered sensorium (93.7%), neck stiffness (75%), and vomiting (62.5%). Overall, Cluster of differentiation 4 (CD4) T-lymphocytes count was <100 cells/µl except 1 case in which the CD4 T-lymphocytes count was 137 cells/µl. No concomitant cryptococcal and tubercular meningitis case was detected. All 16 patients responded initially to induction therapy of IV amphotericin B 1 mg/kg and fluconazole 800 mg daily for 2 weeks. Subsequently, 4 (25%) patients were lost for follow-up and 2 (12.5%) patients expired during their hospital stay. CONCLUSION: As the clinical and radiological pictures of CM are often non-pointing, routine mycological evaluation is necessary for early definite diagnosis and subsequent initiation of appropriate therapy as the majority of patients respond well to treatment if started early.

Urinary Tract Infection due to Paenibacillus alvei in a Chronic Kidney Disease: A Rare Case Report.

Paenibacilli, the Gram positive, aerobic spore bearing bacilli are found normally in the environment. Though these organisms were not known to cause human disease, until recently; few species of this genus have been reported to cause infections in humans. We report here, a case of urinary tract infection in a 60-year-old chronic kidney disease patient due to this rare bacterium. The patient presented with complains of fever, dysuria, and flank pain. Routine and microscopic examination of urine revealed no abnormality except plenty of pus cells and albumin (1+). Bacterial culture showed significant bacteruria and the isolated bacteria was identified to be Paenibacillus alvei based on standard biochemical reactions.

Intestinal protozoans in adults with diarrhea.

BACKGROUND: Diarrhea is one of the most common presenting complaints in human immunodeficiency virus-infected individuals. AIMS: The study was designed to determine the magnitude of opportunistic and nonopportunistic intestinal parasitic infections among diarrheal patients and association between CD4(+) T-cell counts and human immunodeficiency virus (HIV)-infected intestinal parasites. MATERIALS AND METHODS: A cross-sectional study was conducted among 207 enrolled diarrheal patients attending HIV integrated counseling and testing center from January 2012 to December 2012. Stool samples were subjected to special modified Ziehl-Neelsen and chromotrope staining method for detection of opportunistic protozoans. Blood samples were also collected from all study subjects for HIV testing and CD4(+) T-cell counts were estimated by only in HIV-infected patients. RESULTS: Intestinal parasitic pathogens were detected in 46.1% HIV-infected patients and the major pathogens were opportunistic protozoans 32.2% (37/115), most common being Isospora belli 16.5% (19/115) followed by Cryptosporidium parvum 12.2% (14/115). In HIV noninfected diarrheal patients, major pathogens detected were Entamoeba histolytica/Entamoeba dispar 8.7% (8/92) and Ascaris lumbricoides 3.3% (3/92). CONCLUSIONS: The opportunistic intestinal protozoans especially I. belli and C. parvum were most commonly isolated in HIV-infected patients with diarrhea. Majority of the infections occurred in patients when a CD4(+) T-cell counts were less than 200 cells/µl.

Antimicrobial resistance in pathogens causing urinary tract infections in a rural community of Odisha, India.

BACKGROUND: Antimicrobial resistance of urinary tract pathogens has increased worldwide. Empiric treatment of community-acquired urinary tract infection (CA-UTI) is determined by antimicrobial resistance patterns of uropathogens in a population of specific geographical location. OBJECTIVES: This study was conducted to determine the prevalence of CA-UTI in rural Odisha, India, and the effect of gender and age on its prevalence as well as etiologic agents and the resistance profile of the bacterial isolates. MATERIALS AND METHODS: Consecutive clean-catch mid-stream urine samples were collected from 1670 adult patients. The urine samples were processed and microbial isolates were identified by conventional methods. Antimicrobial susceptibility testing was performed on all bacterial isolates by Kirby Bauer's disc diffusion method. RESULTS: The prevalence of UTI was significantly higher in females compared with males (females 45.2%, males 18.4%, OR = 2.041, 95% CI = 1.64-2.52, P ≤ 0.0001). Young females within the age group of 18-37 years and elderly males (≥68 years) showed high prevalence of UTI. Escherichia coli (68.8%) was the most prevalent isolate followed by Enterococcus spp. (9.7%). Amikacin and nitrofurantoin were the most active antimicrobial agents which showed low resistance rate of 5.8% and 9.8%, respectively. CONCLUSION: Our study revealed E. coli as the pre-dominant bacterial pathogen. Nitrofurantoin should be used as empirical therapy for uncomplicated CA-UTIs. In the Indian setting, routine urine cultures may be advisable, since treatment failure is likely to occur with commonly used antimicrobials. Therefore, development of regional surveillance programs is necessary for implementation of national CA-UTI guidelines.

Frequency, risk factors, and antibiogram of Acinetobacter species isolated from various clinical samples in a tertiary care hospital in Odisha, India.

BACKGROUND: For the past two decades, Acinetobacter spp. have emerged as an important pathogen globally in various infections. OBJECTIVES: This study was conducted to determine the frequency, risk factors, and antibiotic resistance pattern of Acinetobacter spp. from various clinical samples. MATERIALS AND METHODS: This retrospective, hospital record-based, cross-sectional study included a total of 8749 clinical samples collected from patients at a tertiary care hospital in Odisha, India from July 2010 to December 2012. The samples were processed and identified by standard protocol. The Acinetobacter isolates were tested for antibiotic resistance by Kirby-Bauer disk diffusion method [according to the Clinical and Laboratory Standards Institute (CLSI) guidelines]. RESULTS: From 8749 clinical samples, 4589 (52.5%) yielded significant growth and only 137 (3%, 137/4589) Acinetobacter spp. were isolated. Maximum (56.9%) isolates were obtained from pus/swab, followed by blood (13.1%) and urine (12.4%). Elderly age, being inpatients, longer duration of stay in the hospital, associated co-morbidity, and invasive procedure were found to be significant risk factors in the setup investigated (P is less than 0.05). Out of 137 isolates, 75 (54.7%) were resistant to more than three classes of antibiotics (multidrug resistant) and 8 (5.8%) were resistant to all commonly used antibiotics (pan-drug resistant). Majority of the isolates were sensitive to imipenem, meropenem, and piperacillin/tazobactam, and showed resistance rates of 19%, 22%, and 23%, respectively. All eight pan-drug resistant isolates were 100% sensitive to colistin. CONCLUSION: This hospital-based epidemiological data will help to implement better infection control strategies and improve the knowledge of antibiotic resistance patterns in our region.

Seroprevalence of chikungunya in southern odisha.

BACKGROUND: The emergence of chikungunya (CHIK) infection was observed in Odisha, India in 2006. Thereafter many cases with symptoms suggestive of CHIK were reported from different districts of Southern-Odisha. This study was aimed to know the seroprevalence, clinical presentations and seasonal trends of CHIK infection in this region. MATERIALS AND METHODS: This study was conducted in a tertiary hospital of this region. Serum samples received in the Department of Microbiology from various districts of Southern-Odisha from April 2011 to March 2012 were included in the study. The samples were tested for CHIK and dengue Immunoglobin M (IgM) antibodies by enzyme-linked immunosorbent assay and malaria parasite by immunochromatographic test (ICT) method. RESULTS: Out of the 678 serum samples tested, 174 were positive for CHIK, 15 for dengue and two samples were positive for both CHIK and dengue IgM antibodies. The most affected age group was 16-45 years. Females were more affected than males. CONCLUSION: The seroprevalence of CHIK among the suspected cases was 25.7%. Co-infection with CHIK and dengue was found to be 1.15%. The infection had spread to new areas during this outbreak.

Prevalence of isosporiasis in relation to CD4 cell counts among HIV-infected patients with diarrhea in Odisha, India.

BACKGROUND: To determine the prevalence of Isospora belli and its correlation with CD4+ cell counts in HIV-positive patients with diarrhea in this region. MATERIALS AND METHODS: Stool samples from 250 HIV-positive patients, including 200 with diarrhea and 50 without diarrhea included in the study were examined for the presence of enteric parasites under microscopy. Prevalence of the enteric parasites with special reference to I. belli in HIV-positive patients with and without diarrhea were calculated and correlated with their CD4+ cell counts. RESULTS: Enteric parasites were detected in 39% of the HIV patients with diarrhea compared to 30% without diarrhea. I. belli was detected in 22% of the patients with diarrhea and in 4% without diarrhea (P = 0.0019). I. belli was the most common parasite, followed by Entamoeba histolytica/dispar (8%) and Cryptosporidium parvum (5%) in HIV-positive patients with diarrhea. In HIV-positive patients without diarrhea, the most common parasite detected was E. histolytica/dispar (12%) followed by C. parvum (6%) and I. belli (4%). The mean CD4 cell count of HIV-positive patients with diarrhea suffering from isosporiasis was 138.35 ± 70.71. In patients with CD4 cell counts <200/µl, I. belli was seen in 36/123 stool samples and 2/27 stool samples which was statistically significant (P = 0.0157). CONCLUSION: I. belli was the predominant parasite with a prevalence of 22% among HIV-positive patients with diarrhea, majority having CD4 cell count <200/µl. This study highlights the importance of routine screening for coccidian parasites in HIV-positive patients with and without diarrhea especially in those with low CD4 cell counts.

New Delhi metallo-beta-lactamase: a weapon for the newly emerging drug-resistant bacteria.

The world has seen the emergence of many micro-organisms in the recent past, which can curb human population with their newly built genetic make-up. The latest addition to this list of panic creating organisms is, bacteria encoding the gene for New Delhi metallo-beta-lactamase (NDM-1). NDM-1 is an enzyme that can hydrolyze and inactivate carbapenems, which are used as a last resort for the treatment of multi-resistant bacterial infections. Names of these bacteria were not found in the medical literature before December 2009, because of which it can take the credit of becoming a powerful emerging bacteria, which are difficult to treat. Besides Escherichia coli and Klebsiella pneumoniae, other bacterial strains have also expressed the gene for NDM-1, which are detected in many countries.

Laboratory diagnosis of chikungunya virus: do we really need it?

Chikungunya (CHIK) fever is a re-emerging Aedes mosquito-transmitted viral disease caused by CHIK virus belonging to the Togaviridae family of genus Alphavirus. The disease is almost self-limiting, occurs with characteristic triad of sudden onset fever, rash and arthritis. During the recent outbreak CHIKV was also found to cause long-term arthralgia, severe neurological disease and even fatalities. Although there are no antiviral or vaccines available for CHIKV, still there are several advantages to diagnose the infection. The present article provides an overview of various diagnostic modalities available and its significance by searching PubMed MeSH terms "Chikungunya virus" and "Diagnosis" for recent articles. The gold standard of CHIKV diagnosis is culture, yet requires facilities and skills. Highly sensitive and specific PCR assays for CHIKV have been developed, but the reagents and equipment are costly for widespread use. Serological diagnosis by detecting IgM antibody is most widely used as it is relatively cheaper and easier to perform. Disadvantages of antibody testing are cross-reactivity with other alpha viruses, cannot differentiate between recent past and acute infection, and its sensitivity varies in clinical settings. When tested for diagnosing acute CHIKV disease, sensitivities were just 4 to 22% and after 1 week rose to more than 80%. As most acutely infected patients seek medical attention within the first few days of illness, the ideal test should detect RNA or antigen. Therefore, the more realistic aim would be to develop a reliable antigen detection assay that could be used in rural areas, where CHIKV infection often occurs.