Shelter Medicine Sustainability from an Academic Perspective: Challenges and Issues.

A meeting of veterinary school faculty and partners, many associated with shelter medicine, and/or community medicine programming, was convened at the 2019 Shelter Medicine Veterinary Educators Conference in Pullman, WA to discuss challenges with shelter medicine program sustainability and defining the future. The discussion was facilitated by an outside consultant and is summarized in this manuscript. The goal of the meeting was to identify challenges and issues concerning the needs and goals for shelter medicine curricula to have long-term success in academic training. Four themes were identified in the transcripts including external pressure from leadership and other stakeholders, funder expectations, time horizons, and perceptions of shelters and shelter veterinarians. Addressing these challenges will be critical to ensuring stability in academic training in shelter medicine, a critical tool for both learning outcomes for general graduates and specific for veterinarians pursuing shelter medicine as a career.

"I couldn't even talk to the patient": Barriers to communicating with cancer patients as perceived by nursing students.

Communication is closely related to safe practice and patient outcomes. Given that most clinicians fall into routines when communicating with patients, it is important to address communication issues early. This study explores Taiwanese nursing students' experiences of communication with patients with cancer and their families. Senior nursing students who had cared for cancer patients were recruited to participate in focus group interviews. These semi-structured interviews were recorded and transcribed for content analysis. Among the 45 participants, about 36% of them never received any communication training. Up to 76% of the participants stated that their communication with cancer patients was difficult and caused them emotional stress. Subsequent data analysis revealed four themes: disengagement, reluctance, regression and transition. Students' negative communication experiences were related to the patients' terminally ill situation; the students' lack of training, low self-efficacy and power status, poor emotional regulation, and cultural considerations. The findings of this study provide a deeper understanding of nursing students' communication experiences in oncology settings within the cultural context. Early and appropriate communication training is necessary to help students regulate their emotions and establish effective communication skills. Further studies are needed to examine the relationship among students' emotional labour, communication skills and outcomes.

PPARß/δ directs the therapeutic potential of mesenchymal stem cells in arthritis.

OBJECTIVES: To define how peroxisome proliferator-activated receptor (PPAR) ß/δ expression level in mesenchymal stem cells (MSCs) could predict and direct both their immunosuppressive and therapeutic properties. PPARß/δ interacts with factors such as nuclear factor-kappa B (NF-κB) and regulates the expression of molecules including vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1. Since these molecules are critical for MSC function, we investigated the role of PPARß/δ on MSC immunosuppressive properties. METHODS: We either treated human MSCs (hMSCs) with the irreversible PPARß/δ antagonist (GSK3787) or derived MSCs from mice deficient for PPARß/δ (PPARß/δ-/- MSCs). We used the collagen-induced arthritis (CIA) as model of immune-mediated disorder and the MSC-immune cell coculture assays. RESULTS: Modulation of PPARß/δ expression in hMSCs either using GSK3787 or hMSCs from different origin reveals that MSC immunosuppressive potential is inversely correlated with Ppard expression. This was consistent with the higher capacity of PPARß/δ-/- MSCs to inhibit both the proliferation of T lymphocytes, in vitro, and arthritic development and progression in CIA compared with PPARß/δ+/+ MSCs. When primed with proinflammatory cytokines to exhibit an immunoregulatory phenotype, PPARß/δ-/- MSCs expressed a higher level of mediators of MSC immunosuppression including VCAM-1, ICAM-1 and nitric oxide (NO) than PPARß/δ+/+ MSCs. The enhanced NO2 production by PPARß/δ-/- MSCs was due to the increased retention of NF-κB p65 subunit on the κB elements of the inducible nitric oxide synthase promoter resulting from PPARß/δ silencing. CONCLUSIONS: Our study is the first to show that the inhibition or knockdown of PPARß/δ in MSCs primes their immunoregulatory functions. Thus, the regulation of PPARß/δ expression provides a new strategy to generate therapeutic MSCs with a stable regulatory phenotype.

Presentations to an acute medical unit due to headache: a review of 306 consecutive cases.

The United Kingdom National Health Service has recently prioritised the need for ambulatory care pathways for acute headache. The present study sought to better characterise patients referred to an Acute Medical Unit so as to inform pathway development. In 2011, York Hospital received 306 referrals due to acute headache, representing 3% of acute medical admissions. Investigations included CT scan (38%), lumbar puncture (38%), and MRI (18%); there were no specialised investigations in 26%, and 18% of patients were discharged on the day of presentation. Subarachnoid haemorrhage occurred in only 4 patients (1%), meningitis in 10 (3%), and intracranial tumour in 5 (2%). The findings indicate that a significant proportion of patients with acute headache could be managed by ambulatory care.

Animal shelter management of feline leukemia virus and feline immunodeficiency virus infections in cats.

Approximately 5% of cats in animal shelters in the United States test positive for either feline leukemia virus (FeLV) or feline immunodeficiency virus (FIV), which translates to more than 100,000 positive cats managed by shelters each year. Little is known about the current status of retroviral management in animal shelters, particularly in regions burdened by chronic pet overpopulation and high shelter admissions, such as the southern United States. The purpose of this study was to describe feline retroviral management in Florida shelters. Shelters were surveyed on practices including selection of cats for testing, diagnostic techniques, and outcome options for cats with positive test results. Responses were received from 139 of 153 animal shelters known to admit cats, including 55 municipal shelters (40%), 70 private shelters (50%), and 14 private shelters with municipal contracts (10%). A total of 115 shelters (83%) performed at least some testing, most using combination point-of-care devices for simultaneous FeLV antigen and FIV antibody screening. Of shelters that performed any testing, 56 (49%) tested all cats for FeLV and 52 (45%) tested all cats for both FeLV and FIV. The most common reason for testing was screening adoptable cats (108 shelters; 94%) and cats available for transfer to other organizations (78; 68%). Testing cats in trap-neuter-return/return-to-field programs was least common (21; 18%). Most common outcome options for positive cats included adoption (74; 64%), transfer (62; 54%), and euthanasia (49; 43%). Euthanasia following a positive test result was more common for cats with FeLV (49; 43%) than for cats with FIV (29; 25%) and was more common in municipal shelters, rural shelters, shelters taking in <500 cats a year, and shelters with overall live outcome rates for cats <70%. Although Florida shelter compliance with national guidelines for identification and management of FeLV and FIV positive cats was variable, most had live outcome options for at least some of their cats with positive test results. Increased access to training and practical programmatic tools may help more shelters implement cost-effective testing protocols, reduce risk for transmission to other cats, and support the best outcomes for this vulnerable population of cats.

Effects of differential supplementation of fatty acids during the peripartum and breeding periods of Holstein cows: II. Neutrophil fatty acids and function, and acute phase proteins.

The objectives were to evaluate the effects of differential supplementation of Ca salts (CS) of fatty acids (FA) on plasma acute phase proteins and both FA composition and function (i.e., activity and cytokine production) of neutrophils, during the peripartum and breeding periods. Holstein cows were assigned randomly to receive either CS of palm (PO) or safflower (SO) oils from 30 d prepartum until 35 d postpartum (dpp) and CS of PO or fish oil (FO) from 35 to 160 dpp. Supplementation of CS of FA was at 1.5% of dietary dry matter. Cows (n=32) were sampled three times weekly from parturition to 35 dpp for analyses of plasma concentrations of haptoglobin and fibrinogen. Cows (n=47) were sampled for neutrophil phagocytic and oxidative burst activities toward Escherichia coli and Staphylococcus aureus, and neutrophil abundances of L-selectin and ß(2)-integrin assessed by flow cytometry at 32 d prepartum, within 7h after parturition, and 4 and 7 dpp. Profiles of FA in neutrophils and cytokine production (i.e., tumor necrosis factor alpha, TNF-α, and IL-1ß) were assessed prepartum (n=14), 35 (PO vs. SO; n=26) and 85 dpp (PO vs. FO; n=28). Plasma concentrations of haptoglobin and fibrinogen were greater for cows fed SO compared with PO. The percentage of neutrophils with phagocytic and oxidative burst activities was not affected by transition diets, but activities per neutrophil were greater in SO compared with PO diets at 4 (phagocytosis and oxidative burst) and 7 dpp (oxidative burst). Neutrophil abundance of L-selectin, but not ß(2)-integrin, was greater in SO compared with PO at 4 and 7 dpp. Neutrophil productions of TNF-α and IL-1ß were increased at 35 dpp in SO compared with PO diets, but production of TNF-α was attenuated in FO compared with PO at 85 dpp. Neutrophil ratios of n-6:n-3 FA were greater at 35 dpp in the SO diet and less at 85 dpp in FO compared with PO diets. In conclusion, cows supplemented with CS of SO had improved innate immunity (i.e., acute phase response and neutrophil function) to better cope with the bacterial challenges in the postpartum period. Conversely, CS of FO attenuated neutrophil cytokine production.

Canine H3N8 influenza virus infection in dogs and mice.

An H3N8 influenza virus closely related to equine influenza virus was identified in racing greyhound dogs with respiratory disease in 2004 and subsequently identified in shelter and pet dogs. Pathologic findings in dogs spontaneously infected with canine influenza virus were compared with lesions induced in beagle and mongrel dogs following experimental inoculation with influenza A/canine/Florida/43/2004. BALB/c mice were inoculated with canine influenza virus to assess their suitability as an experimental model for viral pathogenesis studies. All dogs inoculated with virus developed necrotizing and hyperplastic tracheitis and bronchitis with involvement of submucosal glands as well as mild bronchiolitis and pneumonia. Viral antigen was identified in bronchial and tracheal epithelial cells of all dogs and in alveolar macrophages of several dogs. Many dogs that were spontaneously infected with virus also developed bacterial pneumonia, and greyhound dogs with fatal spontaneous infection developed severe pulmonary hemorrhage with hemothorax. Virus-inoculated BALB/c mice developed tracheitis, bronchitis, bronchiolitis, and mild pneumonia in association with viral antigen in airway epithelial cells and in type 2 alveolar epithelial cells. Virus was not detected in extrarespiratory sites in any animals. The results indicate that canine influenza virus infection consistently induces acute tracheitis and bronchitis in dogs. Mice may be a useful model for some pathogenesis studies on canine influenza virus infection.

A diagnostic algorithm for detection of antibodies to influenza A viruses in dogs in Italy (2006-2008).

Since 2004, there have been several reports of Influenza A virus (FLUAV) infection in dogs. Dogs have been infected with equine influenza H3N8, avian influenza H3N2 and H5N1, and the pandemic H1N1 virus. Because of recent avian and equine influenza outbreaks in Italy, the objectives of the present study were to estimate the level of exposure of Italian dogs to influenza A viruses and to assess a diagnostic algorithm for detection of FLUAV exposure in dogs. Sera collected from 6,858 dogs from 2006 to 2008 were screened in a competitive enzyme-linked immunosorbent assay (cELISA) for antibodies to the highly conserved influenza A nucleoprotein. Samples positive in the cELISA were confirmed by testing in hemagglutination inhibition (HI) and fluorescent antibody test (FAT). Two seropositive dogs had antibodies to H3 hemagglutinin proteins, consistent with exposure to recent canine and equine subtype H3N8 viruses. Using a Bayesian model, the sensitivity and specificity of the cELISA were estimated as 93.98% (probability intervals [PI]: 81.67-99.08%) and 98.71% (PI: 98.43-98.96%), respectively. After accounting for the imperfect sensitivity and specificity of the cELISA, the Bayesian posterior prevalence of FLUAV exposure among tested Italian dogs was 0.5% (PI: 0.1-1.4%). The study results indicate that screening with a cELISA for influenza A nucleoprotein antibody, followed by confirmatory testing with HI and/or FAT, is a highly sensitive and highly specific approach for diagnosing FLUAV exposure in dogs.

Diagnostic application of H3N8-specific equine influenza real-time reverse transcription polymerase chain reaction assays for the detection of Canine influenza virus in clinical specimens.

The objective of the current study was to determine the capability of 3 recently described one-step TaqMan real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays targeting the nucleoprotein (NP), matrix (M), and hemagglutinin (HA) genes of H3N8 Equine influenza virus (EIV NP, EIV M, and EIV HA3 assays, respectively) to detect Canine influenza virus (CIV). The assays were initially evaluated with nucleic acid extracted from tissue culture fluid (TCF) containing the A/canine/FL/43/04 strain of Influenza A virus associated with the 2004 canine influenza outbreak in Florida. The EIV NP, EIV M, and EIV HA3 assays could detect CIV nucleic acid at threshold cycle (Ct) values of 16.31, 23.71, and 15.28, respectively. Three assays using TCF or allantoic fluid (AF) samples containing CIV (n  =  13) and archived canine nasal swab samples (n  =  20) originally submitted for laboratory diagnosis of CIV were further evaluated. All TCF and AF samples, together with 10 nasal swab samples that previously tested positive for virus by attempted isolation in embryonated hens' eggs or Madin-Darby canine kidney cells, were positive in all 3 real-time RT-PCR assays. None of the 3 assays detected the H1N1 Swine influenza virus strain in current circulation. These findings demonstrate that previously described real-time RT-PCR assays targeting NP, M, and H3 HA gene segments of H3N8 EIV are also valuable for the diagnosis of CIV infection in dogs. The assays could expedite the detection and identification of CIV.

Prevalence of protective antibody titers for canine distemper virus and canine parvovirus in dogs entering a Florida animal shelter.

OBJECTIVE: To determine the proportion of dogs entering an animal shelter with protective antibody titers (PATs) for canine distemper virus (CDV) and canine parvovirus (CPV) and identify factors associated with having a PAT. DESIGN: Cross-sectional study. ANIMALS: 431 dogs admitted to an open-admission municipal animal shelter in north central Florida with a history of infectious disease outbreaks. PROCEDURES: Blood was collected from dogs on the day of admission to the shelter. Antibody titers for CDV and CPV were measured by virus neutralization and hemagglutination inhibition, respectively. Age, sex, neuter status, address of origin, source (stray or previously owned), health status (healthy or not healthy), and outcome (adoption, euthanasia, or reclaimed by owner) data were also collected. RESULTS: Overall, 64.5% (278/431) of dogs had insufficient titers for antibodies against CDV, CPV, or both. A total of 153 (35.5%) dogs had PATs for both CDV and CPV, 33 (7.7%) had PATs for CDV but not CPV, 136 (31.5%) had PATs for CPV but not CDV, and 109 (25.3%) did not have PATs for either virus. Older dogs were more likely to have PATs for CDV and CPV. Neutered dogs were more likely to have PATs for CDV. Factors not associated with having a PAT included source, health status, and type of community from which the dog originated. CONCLUSIONS AND CLINICAL RELEVANCE: Most dogs had insufficient antibody titers for CDV, CPV, or both at the time of admission to the animal shelter. Findings support current guidelines recommending vaccination of all dogs immediately upon admission to shelters, regardless of source or physical condition.

PPARß/δ-dependent MSC metabolism determines their immunoregulatory properties.

Mesenchymal stem cell (MSC)-based therapy is being increasingly considered a powerful opportunity for several disorders based on MSC immunoregulatory properties. Nonetheless, MSC are versatile and plastic cells that require an efficient control of their features and functions for their optimal use in clinic. Recently, we have shown that PPARß/δ is pivotal for MSC immunoregulatory and therapeutic functions. However, the role of PPARß/δ on MSC metabolic activity and the relevance of PPARß/δ metabolic control on MSC immunosuppressive properties have never been addressed. Here, we demonstrate that PPARß/δ deficiency forces MSC metabolic adaptation increasing their glycolytic activity required for their immunoregulatory functions on Th1 and Th17 cells. Additionally, we show that the inhibition of the mitochondrial production of ATP in MSC expressing PPARß/δ, promotes their metabolic switch towards aerobic glycolysis to stably enhance their immunosuppressive capacities significantly. Altogether, these data demonstrate that PPARß/δ governs the immunoregulatory potential of MSC by dictating their metabolic reprogramming and pave the way for enhancing MSC immunoregulatory properties and counteracting their versatility.

The experiences of spirituality among adults with mental health difficulties: a qualitative systematic review.

AIMS: Despite an increasing awareness of the importance of spirituality in mental health contexts, a 'religiosity gap' exists in the difference in the value placed on spirituality and religion by professionals compared with service users. This may be due to a lack of understanding about the complex ways people connect with spirituality within contemporary society and mental health contexts, and can result in people's spiritual needs being neglected, dismissed or pathologised within clinical practice. The aim of this qualitative systematic review is to characterise the experiences of spirituality among adults with mental health difficulties in published qualitative research. METHODS: An electronic search of seven databases was conducted along with forward and backward citation searching, expert consultation and hand-searching of journals. Thirty-eight studies were included from 4944 reviewed papers. The review protocol was pre-registered (PROSPERO:CRD42017080566). RESULTS: A thematic synthesis identified six key themes: Meaning-making (sub-themes: Multiple explanations; Developmental journey; Destiny v. autonomy), Identity, Service-provision, Talk about it, Interaction with symptoms (sub-themes: Interactive meaning-making; Spiritual disruption) and Coping (sub-themes: Spiritual practices; Spiritual relationship; Spiritual struggles; Preventing suicide), giving the acronym MISTIC. CONCLUSIONS: This qualitative systematic review provides evidence of the significant role spirituality plays in the lives of many people who experience mental health difficulties. It indicates the importance of mental health professionals being aware of and prepared to support the spiritual dimension of people using services. The production of a theory-based framework can inform efforts by health providers to understand and address people's spiritual needs as part of an integrated holistic approach towards care.

Influenza A virus (H3N8) in dogs with respiratory disease, Florida.

In 2004, canine influenza virus subtype H3N8 emerged in greyhounds in the United States. Subsequent serologic evidence indicated virus circulation in dog breeds other than greyhounds, but the virus had not been isolated from affected animals. In 2005, we conducted virologic investigation of 7 nongreyhound dogs that died from respiratory disease in Florida and isolated influenza subtype H3N8 virus. Antigenic and genetic analysis of A/canine/Jacksonville/2005 (H3N8) and A/canine/Miami/2005 (H3N8) found similarity to earlier isolates from greyhounds, which indicates that canine influenza viruses are not restricted to greyhounds. The hemagglutinin contained 5 conserved amino acid differences that distinguish canine from equine lineages. The antigenic homogeneity of the canine viruses suggests that measurable antigenic drift has not yet occurred. Continued surveillance and antigenic analyses should monitor possible emergence of antigenic variants of canine influenza virus.

The relationship between meal frequency and body mass index in black and white adolescent girls: more is less.

OBJECTIVE: To document meal frequency and its relationship to body mass index (BMI) in a longitudinal sample of black and white girls from ages 9-19 years. DESIGN: Ten-year longitudinal observational study. SUBJECTS: At baseline, 1209 Black girls (539 age nine years, 670 age 10 years) and 1,166 White girls (616 age nine years, 550 age 10 years) were enrolled in the National Heart, Lung and Blood Institute Growth and Health Study (NGHS). MEASUREMENTS: Three-day food diaries, measured height and weight and self-reported physical activity and television viewing were obtained at annual in-person visits. RESULTS: Over the course of the study, the percentage of girls eating 3+ meals on all 3 days was reduced by over half (15 vs 6%). Participants who ate 3+ meals on more days had lower BMI-for-age z-scores. Black girls, but not white girls, who ate 3+ meals on more days were less likely to meet criteria for overweight. CONCLUSION: Meal frequency was related to BMI and should be considered when developing guidelines to prevent childhood overweight.

Differentiation of feline immunodeficiency virus vaccination, infection, or vaccination and infection in cats.

BACKGROUND: Serodiagnosis of feline immunodeficiency virus (FIV) is complicated by the use of a formalin-inactivated whole-virus FIV vaccine. Cats respond to immunization with antibodies indistinguishable from those produced during natural infection by currently available diagnostic tests, which are unable to distinguish cats that are vaccinated against FIV, infected with FIV, or both. HYPOTHESIS: An enzyme-linked immunosorbent assay (ELISA) detecting antibodies against formalin-treated FIV whole virus and untreated transmembrane peptide will distinguish uninfected from infected cats, regardless of vaccination status. ANIMALS: Blood samples were evaluated from uninfected unvaccinated cats (n = 73 samples), uninfected FIV-vaccinated cats (n = 89), and FIV-infected cats (n = 102, including 3 from cats that were also vaccinated). METHODS: The true status of each sample was determined by virus isolation. Plasma samples were tested for FIV antibodies by a commercial FIV diagnostic assay and an experimental discriminant ELISA. RESULTS: All samples from uninfected cats were correctly identified by the discriminant ELISA (specificity 100%). Of the samples collected from FIV-infected cats, 99 were correctly identified as FIV-infected (sensitivity 97.1%). CONCLUSIONS AND CLINICAL IMPORTANCE: With the exception of viral isolation, the discriminant ELISA is the most reliable assay for diagnosis of FIV. A practical strategy for the diagnosis of FIV infection would be to use existing commercial FIV antibody assays as screening tests. Negative results with commercial assays are highly reliable predictors for lack of infection. Positive results can be confirmed with the discriminant ELISA. If the discriminant ELISA is negative, the cat is probably vaccinated against FIV but not infected. Positive results are likely to represent infection.

Infectious diseases of dogs and cats on Isabela Island, Galapagos.

BACKGROUND: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans. HYPOTHESIS: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations. ANIMALS: Ninety-five dogs and 52 cats presented during a neutering campaign. METHODS: A prospective cross-sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases. RESULTS: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66-67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia/Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis. CONCLUSIONS AND CLINICAL IMPORTANCE: Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions.

Comparison of intratesticular injection of zinc gluconate versus surgical castration to sterilize male dogs.

OBJECTIVE: To compare castration of dogs by use of intratesticular injection of zinc gluconate with traditional surgical procedures in terms of acceptance by pet owners, ease of use, and short-term outcomes on Isabela Island of the Galápagos Islands. ANIMALS: 161 privately owned male dogs admitted to a neuter program. PROCEDURES: Medical records of male dogs neutered during a 4-week animal control campaign were reviewed to collect information regarding signalment, method of castration, complication rate, and treatment outcomes. RESULTS: Of the 161 dogs admitted for castration, 58 were surgically castrated and 103 were treated with zinc gluconate. Dogs were returned to their owners for observation following castration. Wound dehiscence occurred in 2 skin incisions, representing 3.4% of the 58 dogs that underwent bilateral orchiectomy. Necrotizing zinc-gluconate injection-site reactions occurred in 4 dogs receiving injection volumes near the maximum label dose (0.8 to 1.0 mL), representing 3.9% of the zinc-gluconate procedures. Surgical wound complications were treated by superficial wound debridement and resuturing, in contrast to zinc-gluconate injection-site reactions, which all required orchiectomy and extensive surgical debridement, including scrotal ablation in 2 dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Low cost, ease of use, and cultural acceptance of a castration technique that does not require removal of the testes make zinc gluconate a valuable option for large-scale use in dogs, particularly in remote locations lacking sophisticated clinical facilities or skilled surgeons and staff. Further investigation is needed to identify risk factors in dogs for adverse reactions to zinc gluconate and to develop strategies for avoidance.

Effects of anesthesia and surgery on serologic responses to vaccination in kittens.

OBJECTIVE: To determine the effects of anesthesia and surgery on serologic responses to vaccination in kittens. DESIGN: Prospective controlled trial. ANIMALS: 32 specific-pathogen-free kittens. PROCEDURES: Kittens were assigned to 1 of 4 treatment groups: neutering at 7, 8, or 9 weeks of age or no neutering. All kittens were inoculated with modified-live virus vaccines against feline panleukopenia virus (FPV), feline herpesvirus (FHV), and feline calicivirus (FCV) at 8, 11, and 14 weeks of age and inactivated rabies virus (RV) at 14 weeks of age. Serum antibody titers against FPV, FHV, and FCV were determined at 8, 9, 11, 14, and 17 weeks of age; RV titers were determined at 14 and 17 weeks of age. RESULTS: Serologic responses of kittens neutered at the time of first vaccination (8 weeks) were not different from those of kittens neutered 1 week before (7 weeks) or 1 week after (9 weeks) first vaccination or from those of kittens that were not neutered. In total, 31%, 0%, 69%, and 9% of kittens failed to develop adequate titers against FPV, FCV, FHV, and RV, respectively, by 17 weeks of age. CONCLUSIONS AND CLINICAL RELEVANCE: Neutering at or near the time of first vaccination with a modified-live virus vaccine did not impair antibody responses in kittens. Many kittens that were last vaccinated at 14 weeks of age had inadequate antibody titers at 17 weeks of age. Kittens may be vaccinated in the perioperative period when necessary, and the primary vaccination series should be extended through at least 16 weeks of age.

Rapid Diagnosis of Babesia gibsoni by Point-of-Need Testing by Insulated Isothermal PCR in Dogs at High Risk of Infection.

BACKGROUND: Dogs seized by law enforcement agencies during dogfighting investigations are at increased risk of Babesia gibsoni infection. A rapid and cost-effective diagnostic test would increase the feasibility of mass screening of dogs for infection and monitoring treatment efficacy in B. gibsoni-infected dogs. OBJECTIVE: To determine the performance of a point-of-need insulated isothermal PCR (iiPCR) test for diagnosis of B. gibsoni in dogs rescued in dogfighting investigations. ANIMALS: Two hundred and thirty-three dogs seized in dogfighting investigations. METHODS: Cross-sectional study. Whole blood samples were tested for B. gibsoni and Babesia spp. by iiPCR. Results were compared to a reference standard comprised of concordant results from real-time PCR in a commercial diagnostic laboratory and antibody titers. RESULTS: The iiPCR system was quick to learn, portable, and had a short processing time of <2 hours. Sensitivity and specificity of the iiPCR assay for B. gibsoni were 90% (95% confidence interval [CI] 81-95%) and 99% (CI, 95-100%), respectively. Sensitivity and specificity of the iiPCR assay for Babesia spp. were 87% (CI, 78-93%) and 98% (CI, 0.94-99%), respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: The iiPCR system produced few false-positive results, indicating that positive results are likely to represent true infections when used in high-risk animals. The iiPCR system can fail to identify 10-15% of truly infected dogs. However, the portability, speed, and economy of the iiPCR system compared to testing through a reference laboratory can allow rescue groups to screen and identify infection in more dogs.

Viral gene expression and provirus load of Orf-A defective FIV in lymphoid tissues and lymphocyte subpopulations of neonatal cats during acute and chronic infections.

Neonatal cats were infected with a wild type (JSY3) or orf-A defective (JSY3DeltaORF-A) feline immunodeficiency virus (FIV) to determine the provirus load and level of viral gene expression at the acute versus chronic stages of infection. FIV DNA in the thymus, lymph node, peripheral blood mononuclear cells (PBMCs) and lymphocyte subpopulations at week 8 post-infection was lower in animals infected with JSY3DeltaORF-A as compared to that of JSY3. At week 16 we observed no significant difference in provirus load between the two groups except for B cells where it was higher in the JSY3 infection. In B cells proviral burden was found to be the same in animals infected with JSY3 for both time points. In the chronic stage, therefore, proviral burden dominates in B cells for JSY3, whereas the level of JSY3DeltaORF-A was lower with comparable values for all lymphocytes at both weeks 8 and 16. Gene expression profiles as measured by real time PCR for gag and rev transcripts revealed decreased levels of JSY3DeltaORF-A mRNAs as compared to that of JSY3. The JSY3 chronic phase infection showed viral gene expression to be higher in B cells relative to CD4+ and CD8+ cells. The presence of viral RNA in CD8 and B cells during the chronic infection implicates active virus replication. Hematological profiles revealed that there was a decline in the number of B cells in JSY3DeltaORF-A-infected cats during the chronic stage of infection while no significant change was observed in animals infected with the wild type virus. Comparative analysis of cell numbers to provirus load and levels of viral transcripts in CD4+ and CD8+, however, did not correlate cell numbers to the levels of viral DNA and gene expression. It remains to be determined whether the relatively high virus burden in B cells as compared to CD4+ and CD8+ cells reflects a role for Orf-A in a shift to B cell virus load during the chronic stage of FIV infection.