Zika virus is causally linked with congenital microcephaly and may be associated with pregnancy loss. However, the mechanisms of Zika virus intrauterine transmission and replication and its tropism and persistence in tissues are poorly understood. We tested tissues from 52 case-patients: 8 infants with microcephaly who died and 44 women suspected of being infected with Zika virus during pregnancy. By reverse transcription PCR, tissues from 32 (62%) case-patients (brains from 8 infants with microcephaly and placental/fetal tissues from 24 women) were positive for Zika virus. In situ hybridization localized replicative Zika virus RNA in brains of 7 infants and in placentas of 9 women who had pregnancy losses during the first or second trimester. These findings demonstrate that Zika virus replicates and persists in fetal brains and placentas, providing direct evidence of its association with microcephaly. Tissue-based reverse transcription PCR extends the time frame of Zika virus detection in congenital and pregnancy-associated infections.

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BackgroundHuman adenoviruses (HAdVs) are known causes of respiratory illness outbreaks in congregate settings, but cases and clusters are less well described from community settings in the United States. During December 2016–February 2017, the New Jersey Department of Health received reports of HAdV infections from 3 sources in 3 adjacent counties. We investigated to characterize the epidemiologic, laboratory, and clinical features of this HAdV outbreak.MethodsA case was defined as a New Jersey resident with acute respiratory illness during December 1, 2016–March 31, 2017 with laboratory identification of HAdV genome type 7d (HAdV-7d). Human adenovirus was detected by real-time and conventional polymerase chain reaction and molecular typed by partial hexon capsid protein gene sequencing. The HAdV genome type was identified by whole genome sequencing analysis. Available medical, public health, and surveillance records were reviewed.ResultsWe identified 12 cases, including 3 treatment facility patients, 7 college students, and 2 cases at a tertiary-care hospital. Four cases died; all had underlying comorbidities. Nine HAdV-7d whole genome sequences obtained from all 3 sites were nearly identical.ConclusionsTransmission of HAdV-7d occurred in community and congregate settings across 3 counties and resulted in severe morbidity and mortality in some cases with underlying comorbidities. Clinicians and local and state health departments should consider HAdV in patients with severe respiratory infection.

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In euryhaline teleosts, reorganization of gill tight junctions during salinity acclimation involves dynamic expression of specific claudin (Cldn) paralogs. We identified four transcripts encoding Cldn tight junction proteins in the tilapia gill transcriptome: cldn10c, cldn10e, cldn28a and cldn30. A tissue distribution experiment found cldn10c and cldn10e expression levels in the gill to be 100-fold higher than any other tissues examined. cldn28a and cldn30 levels in the gill were 10-fold greater than levels in other tissues. Expression of these genes in Mozambique tilapia was examined during acclimation to fresh water (FW), seawater (SW), and in response to hormone treatments. Transfer of tilapia from FW to SW elevated cldn10c and cldn10e, while cldn28a and cldn30 were stimulated following transfer from SW to FW. In hypophysectomized tilapia transferred to FW, pituitary extirpation induced reduced expression of cldn10c, cldn10e and cldn28a; these effects were mitigated equally by either prolactin or cortisol replacement. In vitro experiments with gill filaments showed that cortisol stimulated expression of all four cldns examined, suggesting a direct action of cortisol in situ. Our data indicate that elevated cldn10c and cldn10e expression is important during acclimation of tilapia to SW possibly by conferring ion specific paracellular permeability. On the other hand, expression of cldn28a and cldn30 appears to contribute to reorganization of branchial epithelium during FW acclimation. Hormone treatment experiments showed that particular FW- and SW-induced cldns are controlled by cortisol and prolactin.

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BackgroundPrompt and accurate identification and differentiation of Mycobacterium tuberculosis-complex (MTBC) from non-tuberculous mycobacteria (NTM) is crucial for the selection of antimicrobial treatment and appropriate public health response. Diagnosis and characterization of mycobacteria is challenging due to diverse clinical presentations, lack of sensitivity of smear microscopy, and fastidious culture identification. Moreover, because of clinical suspicion of noninfectious conditions, specimens are often not processed for culture and formalin-fixed, paraffin-embedded (FFPE) tissues are the only specimens available. For rapid and accurate identification of Mycobacterium spp. from patient tissues, sensitive and specific molecular assays combined with other tissue-based methods are vital.MethodsWe extracted DNA from FFPE tissues from 931 patients with clinical and histopathological suspicion of mycobacterial infection (received during 2013–2016) and evaluated by multistage, multigene targeted Mycobacterium-genus, complexes-and species-specific PCR assays (targets including 16S rRNA, rpoB, groEL, IS6110, RLEP) and sequencing. Tissues were also examined by acid-fast bacilli (AFB) stains and mycobacteria immunohistochemistry (IHC). Assays to detect mutations associated with drug resistance were performed on MTBC cases.ResultsA Mycobacterium species was detected in 465 (50%) cases by PCR and sequencing. Of these, 380 (82%) were positive by Mycobacterium PCR targeting 16S rRNA. 85 cases (18%), including 9 MTBC, 12 M. avium complex and 3 M. leprae, were positive by other PCRs. Co-infection of MTBC and NTM spp. was detected in 5 cases. Of 465 PCR positive cases, 327 (70%) showed immunostaining and 223 (48%) were AFB-positive. Molecular markers for drug resistance were detected in 9 out of 88 (10%) tested MTBC cases.ConclusionFFPE tissue analysis by multigene targeted PCR assays expands the opportunities for rapid identification of Mycobacterium species, allows differentiation of MTBC from NTM, and helps to detect co-infections. Using multigene targeted PCRs in combination with histopathology and IHC improve the accuracy of diagnosis, particularly in the presence of commensal and environmental pathogens.Disclosures All authors: No reported disclosures.

Canine leproid granuloma (CLG) is a chronic form of dermatitis that has been associated with nontuberculous mycobacterial infections in Africa, Oceania, the Americas, and Europe. We report here a case of CLG associated with a member of the Mycobacterium tuberculosis complex (MTBC), which could be of public health concern. An 8-y-old pet dog developed 0.5–1-cm diameter, raised, firm, nonpruritic, alopecic, painless skin nodules on the external aspects of both pinnae. Histologic examination revealed severe pyogranulomatous dermatitis with intracellular Ziehl-Neelsen–positive bacilli that were immunoreactive by immunohistochemistry using a polyclonal primary antibody that recognizes tuberculous and nontuberculous Mycobacterium species. DNA extracted from formalin-fixed, paraffin-embedded skin sections was tested by a Mycobacterium genus–specific nested PCR assay targeting the 16S rRNA gene. BLAST sequence analysis of 214-bp and 178-bp amplicons showed 99.5% identity with members of the MTBC; however, the agent could not be identified at the species level. Although CLG has been associated traditionally with nontuberculous mycobacterial infections, the role of Mycobacterium spp. within the MTBC as a cause of this condition, and the role of dogs with CLG as possible sources of MTBC to other animals and humans, should not be disregarded given its zoonotic potential.