Modular adjuvant-free pan-HLA-DR-immunotargeting subunit vaccine against SARS-CoV-2 elicits broad sarbecovirus-neutralizing antibody responses.

Subunit vaccines typically require co-administration with an adjuvant to elicit protective immunity, adding development hurdles that can impede rapid pandemic responses. To circumvent the need for adjuvant in a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) subunit vaccine, we engineer a thermostable immunotargeting vaccine (ITV) that leverages the pan-HLA-DR monoclonal antibody 44H10 to deliver the viral spike protein receptor-binding domain (RBD) to antigen-presenting cells. X-ray crystallography shows that 44H10 binds to a conserved epitope on HLA-DR, providing the basis for its broad HLA-DR reactivity. Adjuvant-free ITV immunization in rabbits and ferrets induces robust anti-RBD antibody responses that neutralize SARS-CoV-2 variants of concern and protect recipients from SARS-CoV-2 challenge. We demonstrate that the modular nature of the ITV scaffold with respect to helper T cell epitopes and diverse RBD antigens facilitates broad sarbecovirus neutralization. Our findings support anti-HLA-DR immunotargeting as an effective means to induce strong antibody responses to subunit antigens without requiring an adjuvant.

Determination of the frequency, species distribution and antimicrobial resistance of staphylococci isolated from dogs and their owners in Trinidad.

The close contact between humans and their dogs can lead to the commingling of staphylococci and the exchange of mobile genetic elements encoding antimicrobial resistance. The objectives of this study were to determine the species distribution and antimicrobial resistance patterns of staphylococci colonizing canine pets and their owners in Trinidad. Staphylococci were isolated from canine pets and their owners and identified using MALDI-TOF mass spectrometry. Antimicrobial susceptibilities were determined using the Kirby-Bauer disc diffusion method against seven classes of antimicrobial agents. A total of 440 staphylococci were isolated from 112 canine pets and their owners, 53.4% were from canine pets and 46.6% were from owners. Twenty-four species were detected, of which, most isolates (32.5%) belonged to the Staphylococcus intermedius group (SIG). S. sciuri was the most common species of coagulase-negative staphylococci (CoNS) comprising 22.3% of all isolates. Antimicrobial resistance was highest against commonly used antimicrobials, such as penicillin (51.4%), tetracycline (26.1%) and trimethoprim/sulfamethoxazole (18.6%). These antimicrobials also comprised the most common multidrug resistance (MDR) combination. Overall, 19.1% of isolates displayed multidrug resistance. No methicillin-resistant Staphylococcus aureus (MRSA) isolates were detected. However, methicillin resistance was detected in 13.3% and 15.1% of coagulase-positive staphylococci (CoPS) and the CoNS+CoVS (combined CoNS and coagulase-variable staphylococci) group respectively. The presence of methicillin-resistant staphylococci is worrisome because there is the potential for the transfer of these strains between dogs and humans. These strains may act as a reservoir of resistance genes.

Serosurvey for Infectious Agents Associated with Subfertility and Abortion in Dairy Cattle in Trinidad and Tobago, West Indies.

Despite frequent reports of subfertility and abortion in dairy cattle in Trinidad and Tobago (T&T), little is known about the potential infectious and non-infectious causes. This study set out to investigate possible infectious causes of reproductive problems by measuring the seroprevalence of four of the most significant reproductive pathogens in dairy cattle worldwide: Brucella abortus (B. abortus); Neospora caninum (N. caninum), Bovine Viral Diarrhoea virus (BVDV), and Infectious Bovine Rhinotracheitis virus (IBRV). These four reproductive pathogens have been suspected to be present in dairy cattle in T&T for some time but, previously, studies have not been carried out to confirm their presence. Bulk milk samples were collected from 92 dairy farms across Trinidad, representing a total of 1177 dairy cattle. Four dairy farms were selected for individual milk sampling to assess in-farm seroprevalence levels. Milk samples were tested for antibodies to the four pathogens by commercial ELISA kits. The overall farm seroprevalence was 62% for N. caninium and 23% for IBRV, and no antibodies were detected in any of the bulk milk samples for B. abortus or BVDV. Mixed infections for IBRV and N. caninum were common. Seroprevalence levels were between 8% and 65% for N. caninum and between 3% and 53% IBRV on the four individual farms. These results reveal the presence of IBRV and N. caninum for the first time on the island of Trinidad and importantly reveal no evidence for the circulation of BVDV or B. abortus in dairy cattle in Trinidad.

Serological evidence for eight globally important poultry viruses in Trinidad & Tobago.

Viruses affecting poultry cause significant levels of disease leading to severe economic losses among poultry farmers worldwide. The Americas region continues to be vulnerable to the spread of poultry viruses across the continents and Caribbean island chains. In Trinidad and Tobago (T&T) there is limited information on the viruses circulating in poultry. Many flock are vulnerable to infection and there are occasional outbreaks of disease resulting in high levels of morbidity and mortality. This study aims to identify important viruses of poultry circulating in T&T through a broad-based surveillance approach. Serum samples from 29 layer farms in Trinidad and 14 layer farms in Tobago were collected from the eldest laying hens. Samples were tested from unvaccinated birds for antibodies by enzyme-linked immunosorbent assay (ELISA) against Avian influenza virus (AIV), Infectious bronchitis virus (IBV), Newcastle disease virus (NDV), Infectious laryngotracheitis virus (ILTV), Avian pneumovirus (APV), Infectious bursal disease virus (IBDV), Fowl adenovirus Gp1 (FADV) and Egg drop syndrome virus (EDSV). In Trinidad, the estimated true seroprevalence levels of antibodies were 0% (CI 95%: 0-0%) for AIV, 100% (CI 95%: 97-100%) for IBV, 79.8% (CI 95%: 70.6-86.9%) for NDV, 1% (CI 95%: 0-2.6%) for ILTV, 67.55% (CI 95%: 62.3-72.4%) for APV, 94.93% (CI 95%: 88.0-98.6%) for IBDV, 100% (CI 95%: 99.7-100%) for FADV and 67.8% (CI 95%: 62.4-72.8%) for EDSV. In Tobago, seroprevalence levels were 0% (CI 95%: 0-0%) for AIV, 100% (CI 95%: 95.6-100%) for IBV, 80.5% (CI 95%: 70.1-88.5%) for NDV, 29.9% (CI 95%: 20.8-40.6%) for ILTV, 100% (CI 95%: 97.7-100%) for APV, 97.1% (CI95%: 89.9-100%) for IBDV, 100% (CI 95%: 97.5-100%) for FADV and 100% (CI 95%: 99-100%) for EDSV. The results reveal strong evidence for the circulation of IBV, NDV, APV, IBDV, FADV and EDSV in layer poultry on both islands, as well as ILTV in Tobago.

Molecular Identification of Eimeria Species in Broiler Chickens in Trinidad, West Indies.

Coccidiosis is an intestinal disease of chickens of major economic importance to broiler industries worldwide. Species of coccidia found in chickens include Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox, and Eimeria tenella. In recent years, polymerase chain reaction (PCR) has been developed to provide accurate and rapid identification of the seven known Eimeria species of chickens. The aim of this study was to use species-specific real-time PCR (qPCR) to identify which of the seven Eimeria species are present in Trinidad poultry. Seventeen pooled fecal samples were collected from 6 broiler farms (2-5 pens per farm) across Trinidad. Feces were also collected from birds showing clinical signs of coccidiosis in two live bird markets (pluck shops). qPCR revealed the presence of five species of Eimeria (E. acervulina, E. maxima, E. mitis, E. necatrix, and E. tenella), but not E. brunetti or E. praecox. Mixed infections were detected on all broiler farms, and DNA of two highly pathogenic Eimeria species (E. tenella and E.necatrix) was detected in feces taken from clinically sick birds sampled from the two pluck shops.

Seroprevalence of economically important viral pathogens in swine populations of Trinidad and Tobago, West Indies.

The objective of this study was to evaluate the seroprevalence and identify the strains of swine influenza virus (SwIV), as well as the seroprevalence of porcine parvovirus (PPV), transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine respiratory coronavirus (PRCV), porcine circovirus type 2 (PCV-2), and classical swine fever virus (CSFV) in pigs in Trinidad and Tobago (T&T). Blood samples (309) were randomly collected from pigs at farms throughout T&T. Serum samples were tested for the presence of antibodies to the aforementioned viruses using commercial ELISA kits, and the circulating strains of SwIV were identified by the hemagglutination inhibition test (HIT). Antibodies against SwIV were detected in 114 out of the 309 samples (37%). Out of a total of 26 farms, 14 tested positive for SwIV antibodies. HI testing revealed high titers against the A/sw/Minnesota/593/99 H3N2 strain and the pH1N1 2009 pandemic strain. Antibodies against PPV were detected in 87 out of the 309 samples (28%), with 11 out of 26 farms testing positive for PPV antibodies. Antibodies against PCV-2 were detected in 205 out of the 309 samples tested (66%), with 25 out of the 26 farms testing positive for PCV-2 antibodies. No antibodies were detected in any of the tested pigs to PRRSV, TGEV, PRCV, or CSFV.