Determinants of vaccine coverage and timeliness in a northern Pakistani village.

The incidence of vaccine preventable disease in Pakistan remains high despite a long-standing Expanded Program on Immunization (EPI). We describe vaccine completeness, timeliness and determinants of coverage from a remote rural cohort (2012-2014). Vaccination histories were taken from EPI records. Vaccination was complete if all doses were received according to the EPI schedule and timely if doses were not ≥3 days early or ≥ 28 days late. Three models are presented: a multivariable logistic regression of household demographic and socioeconomic factors associated with complete vaccination, a multivariable mixed effects logistic regression assessing whether or not the vaccine was administered late (versus on-time), and a mixed effects multivariable Poisson regression model analysing the interval (in days) between vaccine doses. Of 959 enrolled children with full vaccination histories, 88.2 and 65.1% were fully vaccinated following either the pentavalent or DPT/HBV schedules if measles was excluded; coverage dropped to 50.0 and 27.1% when both doses of measles were included. Sixty-four (6.7%) were unvaccinated. Coverage and timeliness declined with subsequent doses. Migrating into the village after 1995 (95%CI 1.88 to 5.17) was associated with late vaccination. Being male, having an older father, and having parents with at least some formal education reduced the likelihood of a late dose. The interval between doses was consistent at 5 weeks (compared with the 4 weeks recommended by EPI). None of the socio-demographic variables were related to the likelihood of receiving full coverage. Vaccine coverage in Oshikhandass was higher than national averages. Measles vaccine coverage and timeliness were low; special consideration should be paid to this vaccine. The local vaccination schedule differed from the EPI, but the consistency suggests good local administration.

Systematic Determination of Herpesvirus in Free-Ranging Cetaceans Stranded in the Western Mediterranean: Tissue Tropism and Associated Lesions.

The monitoring of herpesvirus infection provides useful information when assessing marine mammals' health. This paper shows the prevalence of herpesvirus infection (80.85%) in 47 cetaceans stranded on the coast of the Valencian Community, Spain. Of the 966 tissues evaluated, 121 tested positive when employing nested-PCR (12.53%). The largest proportion of herpesvirus-positive tissue samples was in the reproductive system, nervous system, and tegument. Herpesvirus was more prevalent in females, juveniles, and calves. More than half the DNA PCR positive tissues contained herpesvirus RNA, indicating the presence of actively replicating virus. This RNA was most frequently found in neonates. Fourteen unique sequences were identified. Most amplified sequences belonged to the Gammaherpesvirinae subfamily, but a greater variation was found in Alphaherpesvirinae sequences. This is the first report of systematic herpesvirus DNA and RNA determination in free-ranging cetaceans. Nine (19.14%) were infected with cetacean morbillivirus and all of them (100%) were coinfected with herpesvirus. Lesions similar to those caused by herpesvirus in other species were observed, mainly in the skin, upper digestive tract, genitalia, and central nervous system. Other lesions were also attributable to concomitant etiologies or were nonspecific. It is necessary to investigate the possible role of herpesvirus infection in those cases.

Novel cetacean morbillivirus in a rare Fraser's dolphin (Lagenodelphis hosei) stranding from Maui, Hawai'i.

Cetacean morbillivirus (CeMV) is a global threat to cetaceans. We report a novel morbillivirus from a Fraser's dolphin (Lagenodelphis hosei) that stranded in Maui, Hawaii in 2018 that is dissimilar to the beaked whale morbillivirus previously identified from Hawaii and to other CeMV strains. Histopathological findings included intranuclear inclusions in bile duct epithelium, lymphoid depletion, rare syncytial cells and non-suppurative meningitis. Cerebellum and lung tissue homogenates were inoculated onto Vero.DogSLAMtag cells for virus isolation and cytopathic effects were observed, resulting in the formation of multinucleated giant cells (i.e., syncytia). Transmission electron microscopy of infected cell cultures also revealed syncytial cells with intracytoplasmic and intranuclear inclusions of viral nucleocapsids, consistent with the ultrastructure of a morbillivirus. Samples of the cerebellum, lung, liver, spleen and lymph nodes were positive for morbillivirus using a reverse transcription-polymerase chain reaction. The resulting 559 bp L gene sequence had the highest nucleotide identity (77.3%) to porpoise morbillivirus from Northern Ireland and the Netherlands. The resulting 248 bp P gene had the highest nucleotide identity to porpoise morbillivirus in Northern Ireland and the Netherlands and to a stranded Guiana dolphin (Sotalia guianensis) in Brazil (66.9%). As Fraser's dolphins are a pelagic species that infrequently strand, a novel strain of CeMV may be circulating in the central Pacific that could have additional population impacts through transmission to other small island-associated cetacean species.

Risk factors for measles outbreak in Ataq and Habban districts, Shabwah governorate, Yemen, February to May 2018.

BACKGROUND: Recent conflict and war in Yemen lead to collapse of the health system, decrease of immunization coverage and spread of many outbreaks. On May 22, 2018, the surveillance officer in Shabwah governorate reported an increased number of suspected measles. On May 24, 2018, a team from Yemen-Field Epidemiology Training Program was sent to investigate. The aims were to describe the outbreak, determine the risk factors for measles infection and recommend control measures. METHODOLOGY: A descriptive followed by case-control study design (1:2 ratio) were performed. National Measles Surveillance Program case definition and predesigned questionnaire were used to collect data from 73 cases and 146 controls. Attack rate (AR), adjusted odds ratios (aOR) and 95% confidence intervals (95%CI) were calculated. P value < 0.05 was considered as the cut point for significant. Epi info version 7.2 was used. RESULTS: A total of 73 suspected cases were found. Almost 53% were from Habban district, 63% were males and 56% were among age group < 5 years. The overall AR was 82/100,000 population. Measles was significantly associated with contact with case (aOR = 27.3, 95% CI:1.3-551.7), malnourished children aged 6-60 months (aOR = 24.9, 95% CI;1.9-329.6) and unvaccinated children (aOR = 17.2, 95% CI:2.9-100.7). The six collected blood samples found to be positive for measles IgM. CONCLUSIONS: Measles outbreak in Ataq and Habban districts was confirmed. Contact with measles cases, malnutrition and un-vaccination were the potential contributing factors of measles outbreak in Shabwah governorate. An urgent vaccination campaign with health education interventions are highly recommended. Reactivation of the outreach immunization services and strengthening surveillance and response systems are top priority to take place at district and governorate levels.

Development of TaqMan-based real-time RT-PCR assay based on N gene for the quantitative detection of feline morbillivirus.

BACKGROUND: Morbilliviruses are categorized under the family of Paramyxoviridae and have been associated with severe diseases, such as Peste des petits ruminants, canine distemper and measles with evidence of high morbidity and/or could cause major economic loss in production of livestock animals, such as goats and sheep. Feline morbillivirus (FeMV) is one of the members of Morbilliviruses that has been speculated to cause chronic kidney disease in cats even though a definite relationship is still unclear. To date, FeMV has been detected in several continents, such as Asia (Japan, China, Thailand, Malaysia), Europe (Italy, German, Turkey), Africa (South Africa), and South and North America (Brazil, Unites States). This study aims to develop a TaqMan real-time RT-PCR (qRT-PCR) assay targeting the N gene of FeMV in clinical samples to detect early phase of FeMV infection. RESULTS: A specific assay was developed, since no amplification was observed in viral strains from the same family of Paramyxoviridae, such as canine distemper virus (CDV), Newcastle disease virus (NDV), and measles virus (MeV), and other feline viruses, such as feline coronavirus (FCoV) and feline leukemia virus (FeLV). The lower detection limit of the assay was 1.74 × 104 copies/µL with Cq value of 34.32 ± 0.5 based on the cRNA copy number. The coefficient of variations (CV) values calculated for both intra- and inter-assay were low, ranging from 0.34-0.53% and 1.38-2.03%, respectively. In addition, the clinical sample evaluation using this assay showed a higher detection rate, with 25 (35.2%) clinical samples being FeMV-positive compared to 11 (15.5%) using conventional RT-PCR, proving a more sensitive assay compared to the conventional RT-PCR. CONCLUSIONS: The TaqMan-based real-time RT-PCR assay targeting the N gene described in this study is more sensitive, specific, rapid, and reproducible compared to the conventional RT-PCR assay targeting the N gene, which could be used to detect early infection in cats.

Molecular characterization of Feline paramyxovirus and Feline morbillivirus in cats from Brazil.

This study is aimed at detecting Feline paramyxovirus (FPaV) and Feline morbillivirus (FeMV) in 35 urine samples from domestic cats, collected in 2019, with or without clinical signs of uropathies using a reverse transcription-polymerase chain reaction (RT-PCR) followed by semi-nested polymerase chain reaction (SN-PCR) assays to amplify a partial paramyxovirus L gene. Eight (22.9%) out of the 35 urine samples were positive for paramyxoviruses. Sequencing and phylogenetic analyses revealed that three samples were positive for FPaV, four samples were positive for FeMV, and it was not possible to determine which virus was present in one RT-SN-PCR positive urine sample. FPaV strains showed 100% nucleotide (nt) identity with each other and 97% nt identity with a Japanese 163 FPaV strain. The FeMV strains showed 85.9% nt identity with each other; three strains were similar to previously described Brazilian FeMV strains, and one strain clustered in a different branch of the phylogenetic tree together with the first described Chinese FeMV strain. This study provides the first description of FPaV strains in cats from Brazil and provides new information about the molecular characteristics of FPaV and FeMV strains circulating in domestic cats in Brazil.

Detection of severe fever with thrombocytopenia syndrome virus and other viruses in cats via unbiased next-generation sequencing.

We used unbiased next-generation sequencing (NGS) to detect unknown viruses in cats. Serum or plasma samples were obtained from clinically ill cats with suspected acute viral infections. Nucleic acid was extracted from serum or plasma samples to construct a complementary DNA library for NGS. Comprehensive nucleotide sequencing analyses enabled detection of the genomes of various viruses, including the severe fever with thrombocytopenia syndrome virus, feline immunodeficiency virus, feline morbillivirus, parvovirus, and Torque teno felis virus. Our findings indicate that comprehensive nucleotide analyses of serum or plasma samples can be used to detect infections with unknown viruses in cats.

A novel real-time PCR to detect Cetacean morbillivirus in Atlantic cetaceans.

Cetacean morbillivirus (CeMV, family Paramyxoviridae) is a re-emergent pathogen associated with severe epizootic outbreaks causing high mortality among cetaceans worldwide. Recently, CeMV caused an unusual mortality event of Guiana dolphins (Sotalia guianensis) in Brazil. Partial sequence of the viral phosphoprotein (P) gene showed that the Guiana dolphin morbillivirus (GDMV) might represent a new lineage of CeMV. This study aimed to develop a molecular technique to detect the most common CeMV strains known to circulate in the Atlantic Ocean: GDMV, Dolphin morbillivirus (DMV) and Pilot-whale morbillivirus (PWMV). A sensible real-time reverse transcription polymerase chain reaction (RT-qPCR) method based on intercalating dye, targeting the P gene was described. This assay successfully detected GDMV, PWMV and DMV from field samples. Its performance was compared to a RT-qPCR method that specifically detects GDMV. Both assays had high sensibility and excellent intra- and inter-assay reproducibility. A total of 109 field samples from 32 Guiana dolphins were screened for CeMV by conventional RT-PCR in parallel with the RT-qPCR assay. The detection rate increased from 32% to 60% by use of the novel RT-qPCR. The RT-qPCR assay described herein allows rapid and sensitive detection of Atlantic CeMV strains, and is potentially suitable for screening of CeMV globally.

Increased proportion of apoptotic cells in cat kidney tissues infected with feline morbillivirus.

In order to study potential pathogenic mechanisms of feline morbillivirus (FeMV) in infected kidney cells, we performed a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and an immunofluorescence assay (IFA) with an anti-FeMV P protein antibody on a total of 38 cat kidney tissues, 12 of which were positive for FeMV. Among these samples, we detected significantly larger numbers of apoptotic cells in FeMV-positive tissues than in FeMV-negative tissues, and in these tissues, a substantial percentage of TUNEL-positive (TUNEL+) cells contained the FeMV P protein (mean, 37.4; range, 17.4-82.9), suggesting that induction of apoptosis may be an important mechanism for pathological changes associated with FeMV infection in cat kidney tissues.

Molecular epidemiology and genome analysis of feline morbillivirus in household and shelter cats in Thailand.

BACKGROUND: Feline morbillivirus (FeMV) has been discovered in domestic cats associated with tubulointerstitial nephritis, but FeMV is also detected in healthy cats. This research aimed to identify and characterize the FeMV strains detected in a Thai cat population. RESULTS: Two-hundred and ninety-two samples (131 urine and 161 blood) derived from 261 cats (61 sheltered and 200 household cats) were included for investigating the FeMV prevalence using real-time reverse transcription PCR. The overall prevalence of FeMV detection was 11.9% (31/261) among both samples, which accounted for 14.5% (19/131) and 7.5% (12/161) of the urine and blood samples, respectively. Among the FeMV-PCR positive cats, the FeMV-detected prevalence was insignificantly associated with healthy cats (58.1%; 18/31) or urologic cats (41.9%; 13/31). Full-length genome analysis of these FeMV-Thai strains revealed that their genomes clustered together in the FeMV-1A clade with up to 98.5% nucleotide identity. Selective pressure analysis showed that overall FeMV-1 has undergone negative selection, while positive selection sites were more frequently observed in the phosphoprotein gene. CONCLUSIONS: The detected FeMV infections in the Thai cat population were not correlated with urologic disorders, although the virus was more detectable in urine samples. The genetic patterns among the FeMV-1 Thai strains were more consistent. A large-scale study of FeMV in Thai cat samples is needed for further elucidation.

POSTMORTEM COMPUTED TOMOGRAPHY AND MAGNETIC RESONANCE IMAGING FINDINGS IN A CASE OF COINFECTION OF DOLPHIN MORBILLIVIRUS AND ASPERGILLUS FUMIGATUS IN A JUVENILE BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS).

A freshly dead juvenile bottlenose dolphin (Tursiops truncatus), recovered from the waters near Sand Key, Clearwater, FL, was imaged postmortem using computed tomography and magnetic resonance imaging prior to conventional necropsy. The pattern of imaging findings in the brain was compatible with severe multifocal meningoencephalitis with intralesional necrosis and/or hemorrhage, and the pattern of imaging findings in the lungs was compatible with severe multifocal bronchopneumonia. The subsequent investigation included necropsy, histology, culture, and molecular diagnostics and demonstrated disseminated coinfection of dolphin morbillivirus and Aspergillus fumigatus. This is the first report documenting the cross-sectional imaging findings of this important cetacean comorbidity and demonstrates advances in modern, cooperative investigations of marine mammal mortality events.

Feline Morbillivirus, a New Paramyxovirus Possibly Associated with Feline Kidney Disease.

Feline morbillivirus (FeMV) was first isolated in stray cats in Hong Kong in 2012. Since its discovery, the virus has been reported in domestic cats worldwide, including in Hong Kong, Japan, Italy, US, Brazil, Turkey, UK, Germany, and Malaysia. FeMV is classified in the Morbillivirus genus within the Paramyxoviridae family. FeMV research has focused primarily on determining the host range, symptoms, and characteristics of persistent infections in vitro. Importantly, there is a potential association between FeMV infection and feline kidney diseases, such as tubulointerstitial nephritis (TIN) and chronic kidney diseases (CKD), which are known to significantly affect feline health and survival. However, the tropism and viral entry mechanism(s) of FeMV remain unknown. In this review, we summarize the FeMV studies up to date, including the discoveries of various FeMV strains, basic virology, pathogenicity, and disease signs.

Measles and mumps outbreaks in Lebanon: trends and links.

BACKGROUND: Lebanon has experienced several measles and mumps outbreaks in the past 20 years. In this article, a case-based surveillance of both measles and mumps outbreaks in Lebanon was carried out in an attempt to outline factors contributing to the failure of elimination plans and to provide potential solutions. The relationship between the outbreaks of both diseases was described and explored. METHODS: A retrospective descriptive study of confirmed cases of measles and mumps in Lebanon between 2003 and 2018 collected from the Lebanese Ministry of Public Health Epidemiological Surveillance Unit public database was carried out. The information collected was graphically represented taking into consideration dates of reported cases, age groups affected, and vaccination status. RESULTS: The mean number of measles cases was 150.25 cases/year in the 1-4 years age group, 87 cases/year in individuals aging between 5 and 14, and 63.68 cases/year in those > 14 years old. In the latter group, only 18.05% were unvaccinated. The mean number of mumps cases was 30.4 cases/year in the < 4 year age group and 53.8 cases/year in the 10-19 years age group. During the study period, every spike in measles cases was followed by a similar spike in mumps. 9.66% of measles cases occurred in individuals who received at least 2 doses of the vaccine, 52.26% in the unvaccinated, and 38% in those whose vaccination status was undetermined. CONCLUSIONS: Measles in Lebanon is a disease of the pediatric population, but adults remain at risk. Outbreaks of mumps followed those of measles and were mainly among adolescents. Presence of a large number of Syrian refugees in the country may further complicate the situation. Vaccination activities need to be intensified.

Measles outbreak in Dubrovnik-Neretva County, Croatia, May to June 2018.

In May 2018, measles was introduced in the Dubrovnik region by an adult who recently travelled to Kosovo*. Control measures and an outbreak investigation were implemented: 15 epidemiologically-linked cases met the outbreak case definition of a visitor/resident of Dubrovnik-Neretva County with laboratory-confirmed measles and symptom onset beginning on May 19. New cases were identified through hospitals and primary care physicians. Throat swabs, urine and/or serum samples were collected from outbreak cases. RT-PCR detection of viral RNA and IgM/IgG was used to confirm infection. The median age of cases was 33 years, with one 8 month-old infant. Vaccination status was unknown for 9 cases, three were unvaccinated, one case had history of one dose and two cases reported receiving two doses of measles-containing vaccine. There were 11 hospitalisations and one person developed pneumonia. Control teams undertook an extensive search of contacts and implemented a range of control measures. Despite the outbreak occurring at the beginning of the summer tourism season, it was contained and did not spread to neighbouring regions. With continuing measles transmission in Europe, even small outbreaks create a burden on the health system in countries which have eliminated measles, and illustrate the importance of maintaining high immunisation coverage.

Two measles clusters in connection with short inner-European air travels indicating impediments to effective measles control: A cluster analysis.

Importation and transmission of measles via air travel is a public health concern to countries, which are close to or have achieved elimination, i.e., to the majority of countries in Europe. In 2018, two measles cases occurred in Berlin residents, who flew within Europe while being infectious. In addition to contact tracing through passenger manifests, we contacted national authorities in flight destination countries or embarking countries and inquired about epidemiologically linked measles cases to the two Berlin index cases. We identified eight epidemiologically linked cases (six males, median age: 32 years) from three countries associated with three air-travels. Consequently measles was imported to Germany (Bavaria), Denmark and possibly Sweden. Our investigations revealed impediments to an effective public health response indicating the need to revisit current guidelines and methods to better control transmission of measles related to air travel.

You're the Flight Surgeon.

Leary DB. You're the flight surgeon: measles. Aerosp Med Hum Perform. 2019; 90(10):908-910.

Molecular detection and characterisation of feline morbillivirus in domestic cats in Malaysia.

Feline morbillivirus (FeMV), a novel virus from the family of Paramyxoviridae, was first identified in stray cat populations. The objectives of the current study were to (i) determine the molecular prevalence of FeMV in Malaysia; (ii) identify risk factors associated with FeMV infection; and (iii) characterise any FeMV isolates by phylogenetic analyses. Molecular analysis utilising nested RT-PCR assay targeting the L gene of FeMV performed on either urine, blood and/or kidney samples collected from 208 cats in this study revealed 82 (39.4%) positive cats. FeMV-positive samples were obtained from 63/124 (50.8%) urine and 20/25 (80.0%) kidneys while all blood samples were negative for FeMV. In addition, from the 35 cats that had more than one type of samples collected (blood and urine; blood and kidney; blood, urine and kidney), only one cat had FeMV RNA in the urine and kidney samples. Risk factors such as gender, presence of kidney-associated symptoms and cat source were also investigated. Male cats had a higher risk (p = 0.031) of FeMV infection than females. In addition, no significant association (p = 0.083) was observed between the presence of kidney-associated symptoms with FeMV status. From the 82 positive samples, FeMV RNA was detected from 48/82 (58.5%) pet cats and 34/126 (27.0%) shelter cats (p < 0.0001). Partial L and N gene sequencing of the RT-PCR-positive samples showed 85-99% identity to the published FeMV sequences and it was significantly different from all other morbilliviruses. A phylogenetic analysis of the identified Malaysian FeMVs was performed with isolates from Japan, Thailand and China. Molecular characterisation revealed high relatedness of the Malaysian isolates with other Asian FeMVs, indicating that the virus had been circulating only within the region. Therefore, this study confirmed the existence of FeMV among domestic cats in Malaysia. The findings suggest further characterisation of the local isolates, including the whole genome sequencing and that studies at determining the direct consequences of FeMV infection in domestic cats are needed.

Isolation and genome sequences of two Feline Morbillivirus genotype 1 strains from Italy.

Feline morbillivirus (FeMV) is a novel viral paramyxovirus detected in cats. FeMV is suspected to be associated to tubulointerstitial nephritis, but its pathogenic role is far to be clearly understood.  In this short communication, we report the whole genome coding sequences of the first two FeMV strains isolated in Italy.

Feline morbillivirus in Northern Italy: prevalence in urine and kidneys with and without renal disease.

Feline morbillivirus (FeMV) is an emerging virus that was first described in Hong Kong in 2012. Several reports suggested the epidemiological association of FeMV infection with chronic kidney disease (CKD) in cats. The aim of this study was to investigate the presence and the genetic diversity of FeMV as well as the relationship between FeMV infection and CKD in cats from Northern Italy. Urine (n = 81) and kidney samples (n = 27) from 92 cats admitted to the Veterinary Teaching Hospital of the University of Milan between 2014 and 2017 were investigated for FeMV infection. FeMV RNA was detected in one urine sample (1.23%; 95% CI: 0.03-6.68%) and in two kidneys (7.40%; 95% CI: 0.91-24.28%). FeMV RNA was revealed only in urine or kidneys of cats without evidence of CKD. Phylogenetic analysis showed that the three strains clustered with FeMV strains retrieved from public database, forming a distinct sub-cluster of FeMV. The presence of distinct genotypes of FeMV found in this study is in accordance with previous studies demonstrating that FeMV strains are genetically diverse. A clear relationship between the presence of FeMV infection and CKD in the cats from Northern Italy was not observed, confirming recent reports that do not support the hypothesis that FeMV infection is associated with the development of CKD.