High prevalence and phylogenetic analysis of TT-virus infection in Mongolia

A novel DNA virus, TT-virus (TTV), was isolated from a post-transfusion hepatitis patient in Japan. The prevalence of TTV infection was investigated among patients with chronic liver disease and normal alanine aminotransferase (ALT) volunteers as controls in Mongolia. Polymerase chain reaction (PCR) was employed to detect TTV DNA using specific primers derived from open reading frame 1 (ORF1) of the TTV genome. Nucleotide sequences of samples positive for TTV DNA were determined. The sequences were analyzed by a molecular evolutionary method. Fifty (60.2%) hepatitis patients and 12 (42.9%) volunteers were positive for TTV DNA. The serum ALT levels did not differ significantly between patients with single TTV infection and without TTV, HBV and HCV infection. Similarly, the serum ALT levels did not differ significantly between controls with and without TTV infection. Dual infection of TTV with either HBV or HCV did not affect the ALT levels of hepatitis patients. The molecular evolutionary tree showed that TTV was a heterogeneous virus and all strains could be divided into three genotypes in Mongolia. A new genotype was identified that was distinct from those previously reported.     

Incidence,Risk Factors,and Outcomes of Panton-Valentine Leukocidin-Positive Methicillin-Susceptible Staphylococcus aureus Infections in Auckland,New Zealand

Panton-Valentine leukocidin (PVL) has been linked to invasive community-acquired methicillin-resistant Staphylococcus aureus infections. However, the association between disease and PVL-positive methicillin-susceptible Staphylococcus aureus (MSSA) has not been widely reported. We aimed to examine the epidemiology of PVL in clinical MSSA isolates from patients presenting to Auckland City Hospital. Four hundred eleven MSSA clinical isolates and 93 nasal carriage isolates were collected and tested for the presence of the lukSF-PV genes using PCR. The results were examined in light of host and disease factors. Multilocus sequence typing (MLST) was performed on a random subset of isolates to ensure that there was no single PVL-positive MSSA clone responsible for disease in Auckland. The prevalence of the lukSF-PV genes in MSSA isolates associated with disease (124/335; 37%) was not significantly different from the prevalence of the lukSF-PV genes in MSSA nasal carriage isolates (29/93; 31% [P = 0.33]). PVL-positive MSSA isolates in Auckland are genetically diverse and come from a number of different clonal complexes. PVL-positive infections peaked at between 10 and 20 years of age, with a subsequent decline. Pacific ethnicity, age, diagnosis of skin and soft tissue infection (SSTI), community-onset infection, and the need for surgical intervention were found by multivariate analysis to be independently associated with PVL-positive MSSA infection. More than one-third of MSSA infections in our patient population are caused by PVL-positive strains. Those patients with PVL-positive MSSA infection were more likely to be of Pacific ethnicity, be younger in age, have community-onset infection, have SSTI, and need surgical intervention.Staphylococcus aureus is a nasal commensal that can be detected in up to 20 to 30% of the general population, one-third of whom are persistently colonized (28). S. aureus produces a wide variety of virulence factors that contribute to its ability to colonize, invade, and evade the immune system, which includes Panton-Valentine leukocidin (PVL), a bicomponent, pore-forming toxin encoded by two contiguous genes, lukF-PV and lukS-PV. PVL can cause either neutrophil lysis or apoptosis and contributes to tissue necrosis (25). PVL has been linked to skin and soft tissue infections (SSTIs), necrotizing pneumonia, and bone and joint infections in humans (3, 11, 17). Rabbit and human leukocytes are highly sensitive to PVL-mediated leukocytosis (18), and animal studies have shown that PVL causes more-severe disease in dermonecrosis (7, 27), osteomyelitis (6), and necrotizing pneumonia models (B. A. Diep, L. Chan, and P. Tattevin, presented at the 49th Interscience Conference on Antimicrobial Agents and Chemotherapy, San Francisco, CA, 2009).The presence of PVL has been extensively described for methicillin-resistant S. aureus (MRSA), specifically in association with staphylococcal cassette chromosome mec (SCCmec) type IV and also SCCmec type V (4, 25). The epidemiology of PVL-positive methicillin-susceptible S. aureus (MSSA) has not been reported as extensively, and the lukSF-PV genes are not exclusively linked to the presence of the SCCmec element. In the 1950s MSSA ST80 strains, which were associated with outbreaks of SSTI, harbored the lukSF-PV genes (22). There have also been recent reports of PVL-positive MSSA causing clusters of SSTI and necrotizing pneumonia (5, 15).The vast majority of S. aureus strains in New Zealand are methicillin susceptible (MSSA); the prevalence of methicillin-resistant S. aureus (MRSA) remains low, at about 5% (12). New Zealand has a high incidence of S. aureus disease; the incidence of S. aureus bacteremia in the late 1990s was 41 cases per 100,000 adults per year (12). We aimed to examine the prevalence of the lukSF-PV genes in MSSA isolates responsible for disease and asymptomatic nasal carriage, to determine risk factors for infection with PVL-positive MSSA, and to examine the association between PVL and severity of disease.     

Detection and characterisation of swine hepatitis E virus in New Zealand

The objectives of the present study were to establish the presence of hepatitis E virus (HEV) in New Zealand pigs, first by testing for HEV antibody in pig herds throughout New Zealand to measure the herd prevalence, then by attempting to amplify HEV genomic sequences by PCR. Antibody was measured by two independently designed ELISA serology tests. HEV RNA fragments were amplified by RT-PCR of nucleic acid extracted from faeces of 10-12-week-old piglets using primers targeting ORF1, ORF2, and ORF2/3. PCR products were subject to phylogenetic analysis. Antibody to HEV was found throughout New Zealand pig herds as well as in the different age groups within the herds. Twenty herds from 22 tested were positive for HEV antibody (91% herd prevalence). Phylogenetic analysis of the amplified sequences placed this New Zealand strain of HEV closest to the human European strain It-1 (AF 110390) and U.S. swine strain (AF 082843) with 88% and 83% similarity respectively in ORF1. It was concluded that HEV is widely distributed in the New Zealand pig population. Phylogenetic analysis shows that this is a new HEV strain, grouping most closely with the United States/European cluster, which includes HEV strains of both human and swine origin.     

Anaphylactic deaths in Auckland, New Zealand: a review of coronial autopsies from 1985 to 2005

AIMS: To determine the frequency of anaphylactic deaths amongst coronial autopsy cases performed in the greater Auckland region from 1985 to 2005, and review the circumstances of death and autopsy findings. METHODS: A computerised search for anaphylactic deaths was performed using the Forensic Pathology Department database at Auckland City Hospital. Postmortem reports and police reports were reviewed to determine the circumstances of death. Details recorded included basic demographic data, medical history, agent responsible for the allergic reaction, and pathologic findings at autopsy. RESULTS: A total of 18 cases of anaphylactic deaths were identified for the study period, including nine males and nine females, age range 33-76 years, mean 51.9 years. There were 10 reactions to drugs or contrast media (4 anaesthetic agents, 3 antibiotic, 2 IV contrast media, 1 streptokinase), four to bee/wasp venom, two to seafood, and two undetermined. Death occurred within 1 hour of onset of anaphylaxis in 12 cases. Findings at autopsy included non-specific pulmonary oedema and congestion (n = 13), laryngeal oedema (n = 5), cerebral hypoxia (n = 4) and cutaneous oedema (n = 1). Serum tryptase levels were measured in 15 cases, and were significantly elevated (>20 microg/L) in eight cases. CONCLUSION: Anaphylactic reaction is an uncommon cause of sudden death. In many cases, no specific macroscopic or microscopic findings were detected at autopsy. In the presence of a typical clinical history, postmortem measurement of serum tryptase levels can be a useful diagnostic aid.     

HLA-A*24020102L in the UK blood donor population

The low-expression allele HLA-A*24020102L was identified on a likely haplotype bearing B*55, Cw*01 during the investigation of nine cases (four families and five unrelated subjects) of HLA-A*24 lacking the A24 specificity. A search was made of 70,007 HLA-A, HLA-B, and HLA-C DNA-based typed largely northwestern European Caucasoid blood donors on the British Bone Marrow Registry and Welsh Bone Marrow Donor Registry panels for phenotypes possessing A*24, B*55, Cw*01. Fifty three were found, and 12 of these were subsequently shown to possess A*24020102L by sequence-based typing or polymerase chain reaction with sequence-specific primers. This indicated that the likely A*24, B*55, Cw*01 haplotype has a frequency of 0.000378 in the UK and that approximately 22.6% of these will possess A*24020102L. Consequently, HLA-A*24020102L, B*55, Cw*01 is a principal A*24020102L-bearing haplotype, and the minimum carriage and gene frequencies of A*24020102L are 0.017% and 0.000086, respectively, in UK blood donors.     

Cytomegalovirus infection in a volunteer blood donor population.

Among 223 volunteer blood donors who were studied for evidence of cytomegalovirus (CMV) infection, 58 percent had complement-fixing antibody and 59 percent had indirect hemagglutinating antibody to CMV. No virus was isolated from any donor's washed leukocytes or leukocyte-rich plasma in fibroblast monolayer culture. In seven asymptomatic donors (3 percent), CMV was recovered from urine cultures obtained at the time of blood donation. However, at the time of reexamination, viruria was no longer present and serum antibody titers had not changed. In the three patients studied who received blood from three of the cytomegaloviruric donors, serological evidence of CMV infection developed (fourfold or greater indirect hemagglutinating antibody rise), and one recipient also developed cytomegaloviruria; no illnesses was associated with these infections. Further study is needed to establish that the detection of viruria in donors may identify potentially infective blood.     

Clostridium difficile as a cause of healthcare-associated diarrhoea among children in Auckland,New Zealand: clinical and molecular epidemiology

We aimed to determine the incidence of Clostridium difficile infection (CDI), the molecular epidemiology of circulating C. difficile strains and risk factors for CDI among hospitalised children in the Auckland region. A cross-sectional study was undertaken of hospitalised children <15 years of age in two hospitals investigated for healthcare-associated diarrhoea between November 2011 and June 2012. Stool specimens were analysed for the presence of C. difficile using a two-step testing algorithm including polymerase chain reaction (PCR). C. difficile was cultured and PCR ribotyping performed. Demographic data, illness characteristics and risk factors were compared between children with and without CDI. Non-duplicate stool specimens were collected from 320 children with a median age of 1.2 years (range 3 days to 15 years). Forty-six patients (14 %) tested met the definition for CDI. The overall incidence of CDI was 2.0 per 10,000 bed days. The percentage of positive tests among neonates was only 2.6 %. PCR ribotyping showed a range of strains, with ribotype 014 being the most common. Significant risk factors for CDI were treatment with proton pump inhibitors [risk ratio (RR) 1.74, 95 % confidence interval (CI) 1.09–5.59; p?=?0.002], presence of underlying malignancy (RR 2.71, 95 % CI 1.65–4.62; p?=?0.001), receiving chemotherapy (RR 2.70, 95 % CI 1.41–4.83; p?=?0.003) and exposure to antibiotics (RR 1.17, 95 % CI 0.99–1.17; p?=?0.03). C. difficile is an important cause of healthcare-associated diarrhoea in this paediatric population. The notion that neonatal populations will always have high rates of colonisation with C. difficile may not be correct. Several risk factors associated with CDI among adults were also found to be significant.     

Prevalence of seroreactivity among blood donors in rural population

The seroreactivity pattern amongst blood donors in rural population was studied at S.R.T. Rural Medical College and Hospital, Ambajogai (M. S.). The study period was from January 1996 to December 2001. A total number of 12,240 blood donors were screened. The voluntary donation was 36.98% and replacement donors were 63.02%. No professional donor is bled in our blood bank. The HIV seroreactivity among voluntary donors was 1.56% and 2.11% in replacement donors. The HBsAg seroreactivity was 2.78% in voluntary donors and 4.84% in replacement donors. VDRL seroreactivity is 1.12% in replacement donors. No malarial parasite and HCV seroreactive donor was found in our study period. We have found the magnitude of hepatitis to be far more than that of HIV. Hence testing for HCV routinely is mandatory, besides HBsAg.     

Variables affecting T-lymphocyte subsets in a volunteer blood donor population

Inversion of the normal ratio of helper/inducer (Th) to suppressor/cytotoxic (Ts) T lymphocytes is a characteristic finding in symptomatic and presymptomatic patients with acquired immunodeficiency syndrome (AIDS). As an interim measure to prevent transmission of AIDS via blood transfusion, the Stanford Blood Center performed T-lymphocyte subset analysis as a screening procedure. In this report we present results from 8715 consecutive volunteer blood donors, aged 17-77. The mean Th:Ts ratio was 2.00 +/- 0.70 with a significant trend for increasing Th:Ts with increasing age. In all age groups, donors with antibody to cytomegalovirus (CMV) had lower Th:Ts ratios than CMV-seronegative donors. 1.9% of the donors had Th:Ts values less than or equal to 0.85 and blood from these donors was not used for clinical purposes. Compared to the overall donor population, individuals with Th:Ts less than or equal to 0.85 tended to be male, (79 vs 53%) and CMV sero-positive (71 vs 36%); a majority (83%) had a low absolute number of Th cells as well as a low Th:Ts value. Follow-up of donors with low Th:Ts values revealed that some belonged to AIDS high-risk populations, despite denials at the time of donation. One such donor was diagnosed with disseminated Kaposi's sarcoma 8 months after a markedly abnormal T-lymphocyte subset profile was noted during screening. These results suggest that T-lymphocyte subset analysis is capable of identifying at least some blood donors at risk for transmitting AIDS and also point to variable affecting T-cell subsets in apparently healthy blood donors.     

Molecular characterisation of two divergent variants of grapevine leafroll-associated virus 3 in New Zealand

Partial genomic sequences of two divergent grapevine leafroll-associated virus 3 (GLRaV-3) variants, NZ1-B and NZ2, from New Zealand were determined and analysed (11,827 nt and 7,612 nt, respectively). At the nucleotide level, both variants are more than 20 % different from the previously published GLRaV-3 sequences, from phylogenetic groups 1 to 5. Phylogenetic analysis indicated that NZ1-B is a variant of the previously identified divergent NZ-1, while NZ2 is a novel sequence with only 76 % nucleotide sequence identity to GLRaV-3 variants NZ-1, GH11, and GH30. Therefore, NZ2 is a new variant of GLRaV-3. Amino acid sequence analysis of the NZ1-B and NZ2 coat proteins indicated significant substitutions that are predicted to alter the coat protein structure, which potentially leads to the observed reduced immunological reactivity of both variants to the Bioreba anti-GLRaV-3 conjugated monoclonal antibody.     

Incidence and significance of hepatitis B core antibody in a healthy blood donor population

To determine the current incidence of hepatitis B core antibody (anti-HBc) in a healthy blood donor population, 1,893 donors were screened for anti-HBc. Forty-one (2.16%) were found to be initially positive and 35 (1.85%) repeatably positive. Sera from the repeatably positive donors were further screened for hepatitis B surface antibody (anti-HBs), and hepatitis B virus DNA (HBV DNA) by dot hybridisation. The repeatably positive donors were subsequently recalled for further investigation, and their peripheral blood lymphocytes were also screened for HBV DNA by dot hybridisation. Eighteen (51.4%) of the anti-HBc-positive donors were also anti-HBs-positive. HBV DNA was not detected in the serum or the lymphocytes of any of the anti-HBc-positive donors.     

HTLV‐1 and ‐2 in a first‐time blood donor population in Northeastern Brazil: Prevalence,molecular characterization,and evidence of intrafamilial transmission

Independent epidemiology for respective human T‐cell lymphotropic virus (HTLV) types 1 and 2 is little known in blood donors in Brazil, where screening for HTLV‐1/2 is mandatory at blood banks, but no testing to confirm/differentiate these viruses. Therefore, this study aims to assess the prevalence of HTLV‐1 and ‐2 in a first‐time blood donor population in Northeastern Brazil and to carry out molecular characterization of respective isolates. A cross‐sectional study was conducted at the State Blood Bank in Piauí. Samples were screened for anti–HTLV‐1/2 by enzyme immunoassay, and reactive samples were confirmed using a line immunoassay and polymerase chain reaction (PCR). Of 37 306 blood donors, 47 were anti–HTLV‐1/2 reactive by enzyme immunoassay. After confirmed by line immunoassay, 22 were positive for HTLV‐1 (0.59 per 1000; 95% CI: 0.38‐0.87), 14 were positive for HTLV‐2 (0.37 per 1000; 95% CI: 0.21‐0.61), 1 was indeterminate, and the remaining donors were negative. The HTLV‐1 infection was also confirmed by PCR in all anti–HTLV‐1‐positive samples, and sequencing classified these isolates as belonging to the Transcontinental (A) subgroup of the Cosmopolitan (1a) subtype. Of 14 anti–HTLV‐2‐positive samples, 11 were also PCR positive, which belonged to subtype a (HTLV‐2a/c). In addition, 38 family members of 5 HTLV‐1‐ and 3 HTLV‐2‐infected donors were analyzed. Familial transmission of HTLV‐1 and ‐2 was evidenced in 3 families. In conclusion, in Northeastern Brazil, where HTLV‐1 and ‐2 are endemic, counseling blood donor candidates and their families might play a key role in limiting the spread of these viruses.     

Prevalence and intensity of blood and intestinal parasites in a field population of a Mediterranean lizard, Lacerta lepida

We describe the blood and intestinal parasites in the Ocellated lizard, Lacerta lepida, examining the factors that determine the prevalence and intensity of infection of haemogregarines, and the prevalence of coccidia and nematodes. In relation to haemogregarines, no juveniles were detected as being infected, whereas 71.7 % of adults were infected. The prevalence of infection was positively related to the size of the adults. There were no differences between seasons or sexes in the prevalence or intensity of infection in adults. There were no significant differences in the prevalence of infection by nematodes between ages or sexes, nor in relation to the size of adult lizards, but adult lizards excreting coccidian oocysts tend to be smaller. During the mating period, reproductive activities lead to a decrease in the body condition. However, neither the intensity of haemogregarines infection nor the prevalence of intestinal parasites was related to the lizards body condition.