Evaluation of a Revised Point-of-Care Test for the Detection of Feline Leukaemia p27 Antigen and Anti-p15E Antibodies in Cats.

The first point-of-care (PoC) test (v-RetroFel®; modified version 2021) determining the presence of FeLV p27 antigen and FeLV anti-p15E antibodies has become recently commercially available to identify different feline leukaemia virus (FeLV) infection outcomes. This study aimed to assess this PoC test's performance concerning FeLV p27 antigen and FeLV anti-p15E antibody detection. Sensitivity, specificity, positive and negative predictive values (PPV, NPV) were assessed after ten minutes (recommended) and 20 min (prolonged) incubation times. The test results were evaluated as either positive or negative. Serum samples from 934 cats were included, originating from Italy (n = 269), Portugal (n = 240), Germany (n = 318), and France (n = 107). FeLV p27 antigen and anti-p15E antibodies were measured by reference standard ELISAs and compared to the PoC test results. The PoC test was easy to perform and the results easy to interpret. Sensitivity and specificity for FeLV p27 antigen were 82.8% (PPV: 57.8%) and 96.0% (NPV: 98.8%) after both, ten and 20 minues of incubation time. Sensitivity and specificity for anti-p15E antibodies were 31.4% (PPV: 71.6%) and 96.9% (NPV: 85.1%) after ten minutes incubation time; sensitivity was improved by a prolonged incubation time (20 min) to 40.0% (PPV: 76.3%), while specificity remained the same (96.9%, NPV: 86.7%). Despite the improved sensitivity using the prolonged incubation time, lower than ideal sensitivities for both p27 antigen and especially anti-p15E antibodies were found, indicating that the PoC test in its current version needs further improvement prior to application in the field.

Anti-rabies humoral immune response in cats after concurrent vs separate vaccination against rabies and feline leukaemia virus using canarypox-vectored vaccines.

OBJECTIVES: Some expert groups recommend that cats should be vaccinated with non-adjuvanted feline leukaemia virus (FeLV) and rabies vector vaccines, which, in the European Union, are currently not licensed for concurrent use and have to be administered at least 14 days apart (different from the USA) and thus at separate visits, which is associated with more stress for cats and owners. The aim of this study was to assess the anti-rabies antibody response in cats after vaccination against rabies and FeLV at concurrent vs separate (4 weeks apart) visits using two canarypox-vectored vaccines (Purevax Rabies and Purevax FeLV; Boehringer Ingelheim) and to evaluate the occurrence of vaccine-associated adverse events (VAAEs). METHODS: Healthy FeLV antigen-negative client-owned kittens (n = 106) were prospectively included in this randomised study. All kittens received primary vaccinations against rabies (week 0) and FeLV (weeks 4 and 8). After 1 year, the study group (n = 52) received booster vaccinations against rabies and FeLV concurrently at the same visit (weeks 50-52). The control group (n = 54) received booster vaccinations against rabies (weeks 50-52) and FeLV (weeks 54-56) separately. Anti-rabies virus antibodies (anti-RAV Ab) were determined by fluorescent antibody virus neutralisation assay at weeks 4, 50-52 and 54-56, and compared between both groups using a Mann-Whitney U-test. RESULTS: Four weeks after the first rabies vaccination, 87/106 (82.1%) kittens had a titre ⩾0.5 IU/ml and 19/106 (17.9%) had a titre <0.5 IU/ml. Four weeks after the 1-year rabies booster, all cats had adequate anti-RAV Ab according to the World Organisation for Animal Health (⩾0.5 IU/ml), and the titres of the study group (median = 14.30 IU/ml) and the control group (median = 21.39 IU/ml) did not differ significantly (P = 0.141). VAAEs were observed in 7/106 (6.6%) cats. CONCLUSIONS AND RELEVANCE: Concurrent administration of Purevax FeLV and Purevax Rabies vector vaccines at the 1-year booster does not interfere with the development of anti-RAV Ab or cause more adverse effects and thus represents a better option than separate vaccination visits for cats and owners.

[Options for treatment of feline infectious peritonitis - previously and today]. / Optionen zur Therapie der felinen infektiösen Peritonitis ­ früher und heute.

Feline infectious peritonitis (FIP) is one of the most common infectious diseases in cats that is fatal when untreated. So far, there is no legally available effective treatment in Germany. Treatment options include only symptomatic treatment (e. g. glucocorticoids, propentofylline), immunomodulatory approaches (e. g. interferons, polyprenyl immunostimulant), and antiviral chemotherapy with protease inhibitors (e. g. GC376) or nucleoside analogues (e. g. GS-441524, remdesivir). Symptomatic treatment does not cure FIP but may lead to a short-term improvement of clinical signs in a subset of cats. Immunomodulatory treatment has also not shown to be very promising. In contrary, the antiviral compounds GS-441524 and GC376 exhibited significant efficacy in several studies and their use saved the lives of many cats suffering from FIP. However, both agents are currently not licensed and thus cannot be legally administered by veterinarians in Germany. Legally, cats may only be legally treated with GS-441524 in a few countries (e.g. Great Britain and Australia). In other countries, GS-441524 is imported by cat owners via the black market and administered on their own. This article provides an overview of the available treatment options and an outlook on the legal use of effective antiviral drugs.

Prevalence of Different Courses of Feline Leukaemia Virus Infection in Four European Countries.

Prevalence of progressive feline leukaemia virus (FeLV) infection is known to still be high in cats in Europe, especially in Southern Europe, but the prevalence of other outcomes of FeLV infection has not been determined in most countries. The present study aimed to investigate the prevalence of progressive, regressive, abortive, and focal infection in four European countries, two with a high (Italy, Portugal) and two with a low expected prevalence (Germany, France). Blood samples of 934 cats (Italy: 269; Portugal: 240; France: 107; Germany: 318) were evaluated for the p27 antigen, as well as anti-whole virus, anti-SU, and anti-p15E antibodies by enzyme-linked immunosorbent assay (ELISA) in serum and for proviral DNA by quantitative polymerase chain reaction (qPCR) in whole blood. Positive p27 antigen ELISA results were confirmed by reverse transcriptase-qPCR (RT-qPCR) detecting viral RNA in saliva swabs and/or blood. The outcome of FeLV infection was categorised as progressive (antigen-positive, provirus-positive), regressive (antigen-negative, provirus-positive), abortive (antigen- and provirus-negative, antibody-positive), and focal (antigen-positive, provirus-negative) infection. Overall FeLV prevalence was 21.2% in Italy, 20.4% in Portugal, 9.5% in Germany, and 9.3% in France. Prevalence of progressive, regressive, abortive, and focal infection in Italy was 7.8%, 4.5%, 6.3%, and 2.6%; in Portugal 3.8%, 8.3%, 6.7%, and 1.7%; in Germany 1.9%, 1.3%, 3.5%, and 2.8%; in France 1.9%, 3.7%, 2.8%, and 0.9%, respectively. In conclusion, overall FeLV prevalence is still very high, especially in Southern European countries. Therefore, testing, separation of infected cats, and vaccination are still important measures to reduce the risk of FeLV infection.

Long-term follow-up of cats in complete remission after treatment of feline infectious peritonitis with oral GS-441524.

OBJECTIVES: Feline infectious peritonitis (FIP), a common disease in cats caused by feline coronavirus (FCoV), is usually fatal once clinical signs appear. Successful treatment of FIP with oral GS-441524 for 84 days was demonstrated recently by this research group. The aim of this study was to evaluate the long-term outcome in these cats. METHODS: A total of 18 successfully treated cats were followed for up to 1 year after treatment initiation (9 months after completion of the antiviral treatment). Follow-up examinations were performed at 12-week intervals, including physical examination, haematology, serum biochemistry, abdominal and thoracic ultrasound, FCoV ribonucleic acid (RNA) loads in blood and faeces by reverse transciptase-quantitative PCR and anti-FCoV antibody titres by indirect immunofluorescence assay. RESULTS: Follow-up data were available from 18 cats in week 24, from 15 cats in week 36 and from 14 cats in week 48 (after the start of treatment), respectively. Laboratory parameters remained stable after the end of the treatment, with undetectable blood viral loads (in all but one cat on one occasion). Recurrence of faecal FCoV shedding was detected in five cats. In four cats, an intermediate short-term rise in anti-FCoV antibody titres was detected. In total, 12 cats showed abdominal lymphadenomegaly during the follow-up period; four of them continuously during the treatment and follow-up period. Two cats developed mild neurological signs, compatible with feline hyperaesthesia syndrome, in weeks 36 and 48, respectively; however, FCoV RNA remained undetectable in blood and faeces, and no increase in anti-FCoV antibody titres was observed in these two cats, and the signs resolved. CONCLUSIONS AND RELEVANCE: Treatment with GS-441524 proved to be effective against FIP in both the short term as well as the long term, with no confirmed relapse during the 1-year follow-up period. Whether delayed neurological signs could be a long-term adverse effect of the treatment or associated with a 'long FIP syndrome' needs to be further evaluated.

Clinical Follow-Up and Postmortem Findings in a Cat That Was Cured of Feline Infectious Peritonitis with an Oral Antiviral Drug Containing GS-441524.

This is the first report on a clinical follow-up and postmortem examination of a cat that had been cured of feline infectious peritonitis (FIP) with ocular manifestation by successful treatment with an oral multicomponent drug containing GS-441524. The cat was 6 months old when clinical signs (recurrent fever, lethargy, lack of appetite, and fulminant anterior uveitis) appeared. FIP was diagnosed by ocular tissue immunohistochemistry after enucleation of the affected eye. The cat was a participant in a FIP treatment study, which was published recently. However, 240 days after leaving the clinic healthy, and 164 days after the end of the 84 days of treatment, the cured cat died in a road traffic accident. Upon full postmortem examination, including histopathology and immunohistochemistry, there were no residual FIP lesions observed apart from a generalized lymphadenopathy due to massive lymphoid hyperplasia. Neither feline coronavirus (FCoV) RNA nor FCoV antigen were identified by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunohistochemistry, respectively, in any tissues or body fluids, including feces. These results prove that oral treatment with GS-441524 leads to the cure of FIP-associated changes and the elimination of FCoV from all tissues.

[Feline leukemia virus infection - a guide to diagnosis]. / Infektion mit dem felinen Leukämievirus ­ der Weg zur Diagnose.

Feline leukemia virus (FeLV) infection affects cats worldwide. The course of FeLV infection can change and vary over time. The complex pathogenesis, the availability of many different testing methods, and the interpretation of test results are often challenging for veterinarians. Cats with progressive infection (persistently p27 antigen-positive) shed FeLV mainly through saliva and are therefore considered a source of infection for uninfected cats. Diagnosing regressive infection is often challenging, since it usually cannot be detected by commonly used point of care-tests (p27 antigen test) and thus, it often remains undetected. Nevertheless, cats with regressive infection are FeLV carriers (provirus-positive) and when the immune system is suppressed, reactivation of the infection and FeLV-associated clinical signs can occur. Abortively infected cats are never viraemic, do not shed virus, and do not develop clinical signs. Abortive infection can solely be diagnosed via antibodies detection in blood. A new point-of-care test for the identification of antibodies against FeLV p15E antigen has recently been introduced on the European market and is currently being evaluated.

Fecal Feline Coronavirus RNA Shedding and Spike Gene Mutations in Cats with Feline Infectious Peritonitis Treated with GS-441524.

As previously demonstrated by our research group, the oral multicomponent drug Xraphconn® containing GS-441524 was effective at curing otherwise fatal feline infectious peritonitis (FIP) in 18 feline coronavirus (FCoV)-infected cats. The aims of the current study were to investigate, using samples from the same animals as in the previous study, (1) the effect of treatment on fecal viral RNA shedding; (2) the presence of spike gene mutations in different body compartments of these cats; and (3) viral RNA shedding, presence of spike gene mutations, and anti-FCoV antibody titers in samples of 12 companion cats cohabitating with the treated cats. Eleven of the eighteen treated FIP cats (61%) were shedding FCoV RNA in feces within the first three days after treatment initiation, but all of them tested negative by day 6. In one of these cats, fecal shedding reoccurred on day 83. Two cats initially negative in feces were transiently positive 1-4 weeks into the study. The remaining five cats never shed FCoV. Viral RNA loads in feces decreased with time comparable with those in blood and effusion. Specific spike gene mutations linked to systemic FCoV spread were consistently found in blood and effusion from treated FIP cats, but not in feces from treated or companion cats. A new mutation that led to a not yet described amino acid change was identified, indicating that further mutations may be involved in the development of FIP. Eight of the twelve companion cats shed FCoV in feces. All but one of the twelve companion cats had anti-FCoV antibodies. Oral treatment with GS-441524 effectively decreased viral RNA loads in feces, blood, and effusion in cats with FIP. Nonetheless, re-shedding can most likely occur if cats are re-exposed to FCoV by their companion cats.

Curing Cats with Feline Infectious Peritonitis with an Oral Multi-Component Drug Containing GS-441524.

Feline infectious peritonitis (FIP) caused by feline coronavirus (FCoV) is a common dis-ease in cats, fatal if untreated, and no effective treatment is currently legally available. The aim of this study was to evaluate efficacy and toxicity of the multi-component drug Xraphconn® in vitro and as oral treatment in cats with spontaneous FIP by examining survival rate, development of clinical and laboratory parameters, viral loads, anti-FCoV antibodies, and adverse effects. Mass spectrometry and nuclear magnetic resonance identified GS-441524 as an active component of Xraphconn®. Eighteen cats with FIP were prospectively followed up while being treated orally for 84 days. Values of key parameters on each examination day were compared to values before treatment initiation using linear mixed-effect models. Xraphconn® displayed high virucidal activity in cell culture. All cats recovered with dramatic improvement of clinical and laboratory parameters and massive reduction in viral loads within the first few days of treatment without serious adverse effects. Oral treatment with Xraphconn® containing GS-441524 was highly effective for FIP without causing serious adverse effects. This drug is an excellent option for the oral treatment of FIP and should be trialed as potential effective treatment option for other severe coronavirus-associated diseases across species.

Evaluation of a Point-of-Care Test for Pre-Vaccination Testing to Detect Antibodies against Canine Adenoviruses in Dogs.

(1) Background: Antibody testing is commonly used to assess a dog's immune status. For detection of antibodies against canine adenoviruses (CAVs), one point-of-care (POC) test is available. This study assessed the POC test´s performance. (2) Methods: Sera of 198 privately owned dogs and 40 specific pathogen-free (SPF) dogs were included. The reference standard for detection of anti-CAV antibodies was virus neutralization (VN) using CAV-1 and CAV-2 antigens. Specificity, sensitivity, positive predictive value (PPV), negative predictive value (NPV), and overall accuracy (OA) of the POC test were assessed. Specificity was considered most important. (3) Results: Prevalence of CAV-1 neutralizing antibodies (≥10) was 76% (182/238) in all dogs, 92% (182/198) in the subgroup of privately owned dogs, and 0% (0/40) in SPF dogs. Prevalence of CAV-2 neutralizing antibodies (≥10) was 76% (181/238) in all dogs, 91% (181/198) in privately owned dogs, and 0% (0/40) in SPF dogs. Specificity for detection of CAV-1 antibodies was lower (overall dogs, 88%; privately owned dogs, 56%; SPF dogs, 100%) compared with specificity for detection of CAV-2 antibodies (overall dogs, 90%; privately owned dogs, 65%; SPF dogs, 100%). (4) Conclusions: Since false positive results will lead to potentially unprotected dogs not being vaccinated, specificity should be improved to reliably detect anti-CAV antibodies that prevent infectious canine hepatitis in dogs.

Prevalence and Risk Factors of Feline Immunodeficiency Virus and Feline Leukemia Virus Infection in Healthy Cats in Thailand.

Infections with feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) occur worldwide and are among the most important infectious diseases in cats. The aim of the present study was to determine the prevalence of FIV and FeLV infection in healthy outdoor cats in North, Northeast and Central Thailand. So far, a study on retrovirus prevalence of healthy cats in Thailand in a larger geographic area has not been published yet. In addition, risk factors for FIV and FeLV infections were evaluated. Two hundred sixty healthy cats were prospectively recruited. They originated from 13 locations in North, Northeast, and Central Thailand and were presented for either preventive health care and/or neutering. In each cat, a physical examination was performed to confirm health status. FIV and FeLV status was determined using a commercial rapid enzyme-linked immunosorbent assay (ELISA) (SNAP Combo Plus FeLV/FIV, IDEXX). Risk factors were analyzed by binary logistic regression analysis. Samples of 15/260 (5.8%) cats were positive for FIV antibodies, and 11/260 (4.2%) samples were positive for FeLV antigen. One of the 260 (0.4%) cats was positive for both, FIV and FeLV infection. In binary logistic regression analysis, no parameter was associated with a higher risk for FeLV infection. However, cats had a significantly (p = 0.025) higher risk for FIV infection when they were 2 years or older. FIV and FeLV infections occur in healthy cats in North, Northeast and Central Thailand, but prevalence was lower than expected. No risk factors for FeLV infection were detected, but risk for FIV infection increases with age.

Antibody Response to Canine Adenovirus-2 Virus Vaccination in Healthy Adult Dogs.

BACKGROUND: Re-vaccination against canine adenovirus (CAV) is performed in ≤3-year-intervals but their necessity is unknown. The study determined anti-CAV antibodies within 28 days of re-vaccination and factors associated with the absence of antibodies and vaccination response. METHODS: Ninety-seven healthy adult dogs (last vaccination ≥12 months) were re-vaccinated with a modified live CAV-2 vaccine. Anti-CAV antibodies were measured before vaccination (day 0), and after re-vaccination (day 7, 28) by virus neutralization. A ≥4-fold titer increase was defined as vaccination response. Fisher's exact test and multivariate regression analysis were performed to determine factors associated with the absence of antibodies and vaccination response. RESULTS: Totally, 87% of dogs (90/97; 95% CI: 85.61-96.70) had anti-CAV antibodies (≥10) before re-vaccination. Vaccination response was observed in 6% of dogs (6/97; 95% CI: 2.60-13.11). Time since last vaccination (>3-5 years, OR = 9.375, p = 0.020; >5 years, OR= 25.000, p = 0.006) was associated with a lack of antibodies. Dogs from urban areas were more likely to respond to vaccination (p = 0.037). CONCLUSION: Many dogs had anti-CAV pre-vaccination antibodies, even those with an incomplete vaccination series. Most dogs did not respond to re-vaccination. Based on this study, dogs should be re-vaccinated every 3 years or antibodies should be determined.

Detection of feline coronavirus RNA, spike gene mutations, and feline coronavirus antigen in macrophages in aqueous humor of cats in the diagnosis of feline infectious peritonitis.

Uveitis is common in cats, and is often a feature of feline infectious peritonitis (FIP). We evaluated 3 tools for detection of feline coronavirus (FCoV) in aqueous humor: 1) a 7b gene reverse-transcription real-time PCR (7b-RT-rtPCR) assay to detect FCoV RNA, 2) a spike gene mutation RT-rtPCR (S-RT-rtPCR) assay to detect 2 point mutations in the spike gene of FCoV in cats positive by 7b-RT-rtPCR, and 3) immunocytochemistry (ICC) for detection of FCoV antigen in aqueous humor macrophages. We studied 58 cats, including 31 cats with FIP and 27 control cats. FIP was excluded by postmortem examination and negative immunohistochemistry (IHC). Aqueous humor samples obtained postmortem were assessed using 7b-RT-rtPCR in all cats, and positive samples were evaluated with S-RT-rtPCR. ICC evaluation of aqueous humor samples from 36 of the 58 cats was done using an avidin-biotin complex method and monoclonal anti-FCoV IgG 2A. Sensitivity, specificity, and negative and positive predictive values were calculated including 95% CIs. 7b-RT-rtPCR had a specificity of 100.0% (95% CI: 87.2-100.0) and sensitivity of 35.5% (95% CI: 19.2-54.6). Specificity of S-RT-rtPCR could not be determined because there were no FCoV 7b-RT-rtPCR-positive samples in the control group. Sensitivity of S-RT-rtPCR was 12.9% (95% CI 3.6-29.8). Sensitivity and specificity of ICC were 62.5% (95% CI: 40.6-81.2) and 80.0% (95% CI: 44.4-97.5), respectively. The combination of 7b-RT-rtPCR and IHC could be useful in diagnosing FIP; S-RT-rtPCR did not add value; and ICC of aqueous humor samples cannot be recommended for the diagnosis of FIP.

Antibody response to feline herpesvirus-1 vaccination in healthy adult cats.

OBJECTIVES: Vaccination against feline herpesvirus-1 (FHV-1) is recommended for all cats. However, it is unknown how adult healthy cats with different pre-vaccination antibodies respond to FHV-1 vaccination in the field. The aim of the study was to determine the prevalence of neutralising antibodies against FHV-1 in healthy adult cats and the response to FHV-1 vaccination within 28 days of vaccination. METHODS: One hundred and ten cats (⩾1 year of age) that had not received a vaccination within the past 12 months were vaccinated with a combined FHV-1 vaccine. Antibodies against FHV-1 were determined before vaccination (day 0), on day 7 and day 28 by serum neutralisation test. Uni- and multivariate statistical analyses were used to determine factors associated with the presence of pre-vaccination antibodies and response to vaccination. RESULTS: Pre-vaccination neutralising antibody titres (⩾10) were present in 40.9% of cats (45/110; 95% confidence interval [CI] 32.2-50.3); titres were generally low (range 10-640). Antibody response to vaccination (⩾four-fold titre increase) was observed in 8.3% (9/109; 95% CI 4.2-15.1). Cats ⩾2 years of age were more likely to have pre-vaccination neutralising antibodies than cats aged between 1 and 2 years (odds ratio [OR] 24.619; P = 0.005). Cats from breeders were more likely to have pre-vaccination neutralising antibodies than privately owned cats (OR 7.070; P = 0.007). Domestic shorthair cats were more likely to have an at least four-fold titre increase vs purebred cats (OR 11.22; P = 0.027). CONCLUSIONS AND RELEVANCE: Many cats have no detectable neutralising antibodies against FHV-1 despite previous vaccinations and fail to develop a ⩾four-fold titre increase after vaccination. This is likely because older cats and cats with a higher FHV-1 exposure risk are more likely to get infected with FHV-1 and thus to have FHV-1 neutralizing antibodies. Purebred cats more often fail to develop a ⩾four-fold titre increase after vaccination.

A survey of vaccine history in German cats and owners' attitudes to vaccination.

OBJECTIVES: Vaccination is the most important measure for prevention of feline infectious diseases. Cat owner compliance with vaccination recommendations has been investigated in the UK but not in other European countries. The aim of the present study was to determine cat owners' attitudes towards vaccination in cats in Germany, to identify factors that are associated with the vaccination status of their cats and to compare the results with those of the UK survey. METHODS: The survey was conducted using an online questionnaire and was aimed at respondents throughout Germany. Respondents under 16 years of age, cats that were less than 9 weeks old and veterinarians were excluded. A total of 920 questionnaires were evaluated, and information about cats and respondents was assessed with respect to the current vaccination status of the cats using a linear logistic regression model. RESULTS: The majority of cats (77.9%; n = 717) were vaccinated according to current guidelines; only 5.4% (n = 50; 95% confidence interval [CI] 5.00-9.00) of cats had never received a vaccine. Having visited a cattery, a cat show or travelled abroad in the past 12 months (n = 96/773; odds ratio [OR] 6.95; 95% CI 1.65-52.19) had the highest positive impact on the vaccination status of cats. In addition, detailed veterinary advice about vaccination had a positive impact (n = 275/773; OR 2.09; 95% CI 0.67-6.25) on the attitude of owners towards vaccinating their cats. CONCLUSIONS AND RELEVANCE: A history of travelling abroad or visiting cat shows or a cattery, and thus regulatory requirements, had the greatest positive impact on the current vaccination status of the cats. Veterinary consultation on preventive measures, including vaccination, is crucial for protecting the cat population against infectious diseases.

Detection of feline coronavirus mutations in paraffin-embedded tissues in cats with feline infectious peritonitis and controls.

OBJECTIVES: The amino acid substitutions M1058L and S1060A in the spike protein of feline coronavirus (FCoV) have been postulated to be responsible for the development of the pathogenic feline infectious peritonitis virus (FIPV), which causes feline infectious peritonitis (FIP). The aim of the following study was to investigate the presence of mutated virus in tissue samples of cats with and without FIP. METHODS: The study population consisted of 64 cats, 34 of which were diagnosed with FIP and 30 control cats. All cases underwent autopsy, histopathology and immunohistochemistry (IHC) for FCoV. Furthermore, a genotype-discriminating quantitative reverse transcriptase PCR (RT-qPCR) was performed on shavings of paraffin-embedded tissues to discriminate between cats with FIP and controls, and the sensitivity and specificity of this discriminating RT-qPCR were calculated using 95% confidence intervals (CIs). RESULTS: Specificity of genotype-discriminating RT-qPCR was 100.0% (95% CI 88.4-100.0), and sensitivity was 70.6% (95% CI 52.5-84.9). In cats with FIP, 24/34 tested positive for FIPV. In samples of three control cats and in seven cats with FIP, FCoV was found, but genotyping was not possible owing to low FCoV RNA concentrations. Out of the positive samples, 23 showed the amino acid substitution M1058L in the spike protein and none the substitution S1060A. One sample in a cat with FIP revealed a mixed population of non-mutated FCoV and FIPV (mixed genotype). For one sample genotyping was not possible despite high viral load, and two samples were negative for FCoV. CONCLUSIONS AND RELEVANCE: As none of the control animals showed FCoV amino acid substitutions previously demonstrated in cats with FIP, it can be presumed that the substitution M1058L correlates with the presence of FIP. FCoV was detected in low concentration in tissues of control animals, confirming the ability of FCoV to spread systemically. The fact that no negative controls were included in the IHC protocol could potentially lead to an underestimation of the sensitivity of the RT-qPCR.

Antibody response to feline panleukopenia virus vaccination in cats with asymptomatic retrovirus infections: a pilot study.

OBJECTIVES: Currently, there are only a few studies on how immunocompromised cats, such as cats infected with feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV), respond to vaccination. Therefore, this study measured feline panleukopenia virus (FPV) antibodies in retrovirus-infected cats within a period of 28 days after FPV vaccination, and compared the immune response to that of non-infected cats. METHODS: Eight asymptomatic retrovirus-infected cats (four FeLV, four FIV), and non-infected age-matched control cats (n = 67) were vaccinated with a commercial FPV modified live virus (MLV). Pre- and post-vaccination antibody titres were measured by haemagglutination inhibition (HI) on days 0, 7 and 28. An HI titre ⩾1:40 was defined as protective. An adequate response to vaccination was defined as a four-fold titre increase or higher. Comparison of the immune response of retrovirus-infected and non-infected cats was performed. RESULTS: Pre-vaccination FPV antibody titres ⩾1:40 were present in 100% (n = 8/8; 95% confidence interval [CI] 62.8-100) of retrovirus-infected and in 77.6% (n = 52/67; 95% CI 66.2-86.0) of non-infected cats. An adequate response to vaccination (titre increase ⩾four-fold) was seen in 1/8 retrovirus-infected cats (12.5%; 95% CI 0.1-49.2) compared with 22/67 non-infected cats (32.8%; 95% CI 22.8-44.8). In cats with high pre-vaccination titres (⩾1:160), a four-fold titre increase or higher was observed in 1/8 retrovirus infected cats (12.5%; 95% CI 0.1-49.2) compared with 4/42 non-infected cats (9.5%; 95% CI 3.2-22.6). None of the eight retrovirus-infected cats developed illness or vaccination side effects after vaccination with MLV against FPV within the 28 days. There were no significant differences between groups: for pre-vaccination titres; for at least four-fold titre increases following vaccination in either all cats or the cats with high pre-vaccination titres; and concerning adverse effects. CONCLUSIONS AND RELEVANCE: All retrovirus-infected asymptomatic cats had pre-vaccination FPV antibodies indicating protection against panleukopenia. Response of retrovirus-infected cats to vaccination was similar to the response of non-infected cats.

Antibody response to feline panleukopenia virus vaccination in healthy adult cats.

OBJECTIVES: According to prior studies, between 25.0% and 92.8% of adult cats have antibodies against feline panleukopenia virus (FPV) and thus are likely protected against FPV infection. It is, however, unknown how healthy adult cats with different antibody titres react to FPV vaccination in the field. Therefore, the aim of the study was to measure antibody titres in healthy adult cats within a period of 28 days after vaccination against FPV and to evaluate factors that are associated with a lack of adequate response to vaccination. METHODS: One hundred and twelve healthy adult cats were vaccinated with a vaccine against FPV, feline herpesvirus and feline calicivirus. Antibodies against FPV were determined before vaccination (day 0), on day 7 and day 28 after vaccination by haemagglutination inhibition (HI). A HI titre ⩾1:40 was defined as protective. An adequate response to vaccination was defined as a four-fold titre increase. Uni- and multivariate statistical analysis was used to determine factors associated with an adequate response. RESULTS: Pre-vaccination antibody titres of ⩾1:40 were present in 64.3% (72/112; 95% confidence interval [CI] 55.1-72.6). Only 47.3% (53/112; 95% CI 37.8-57.0) of cats had an adequate response to vaccination. Factors associated with an adequate response to vaccination were lack of previous vaccination (odds ratio [OR] 15.58; 95% CI 1.4-179.1; P = 0.035), lack of antibodies (⩾1:40) prior to vaccination (OR 23.10; 95% CI 5.4-98.8; P <0.001) and breed (domestic shorthair cats; OR 7.40; 95% CI 1.4-38.4; P = 0.017). CONCLUSIONS AND RELEVANCE: As none of the cats with high pre-vaccination antibody titres (⩾1:160) had an at least four-fold increase in FPV antibody titres, measurement of antibodies rather than regular revaccinations should be performed. Thus, evaluation of FPV antibody titre in cats with previous vaccinations against FPV are recommended prior to revaccination.

Risk factors of different hemoplasma species infections in cats.

BACKGROUND: Hemoplasma species (spp.) commonly cause infections in cats worldwide. However, data on risk factors for infections are limited. The aim of this study was to determine the prevalence of hemoplasma spp. infections in cats in Southern Germany and to assess risk factors associated with infection. RESULTS: DNA was extracted from blood samples of 479 cats presented to different veterinary hospitals for various reasons. DNA of feline hemoplasmas was amplified by use of a previously reported PCR assay. Direct sequencing was used to confirm all purified amplicons and compared to hemoplasma sequences reported in GenBank. Results were evaluated in relation to the age, sex, housing conditions, feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) status of the cats. The overall hemoplasma prevalence rate was 9.4% (45/479; 95% CI: 7.08-12.36). 'Candidatus Mycoplasma (M.) haemominutum' (Mhm) DNA was amplified from 42 samples, M. haemofelis from 2, and M. haemocanis from 1 sample. There was a significantly higher risk of hemoplasma infection in cats from multi-cat households, in outdoor cats, as well as in cats with FIVinfection and in cats with abortive FeLV infection, but not in cats with progressive or regressive FeLV infection. CONCLUSIONS: Mhm infection is common in cats in Southern Germany. Higher prevalence in multi-cat households and associations with FeLV infection likely reflect the potential for direct transmission amongst cats. Outdoor access, male gender, and FIV infection are additional risk factors that might relate to aggressive interactions and exposure to vectors.

Detection of Leptospira DNA in urine and presence of specific antibodies in outdoor cats in Germany.

Objectives Clinical manifestation of infection with Leptospira species in cats is rare. Nevertheless, cats can develop specific antibodies against the spirochetes after infection. In Canada, Taiwan and the USA it was recently demonstrated that naturally infected cats can also shed DNA from pathogenic Leptospira species in their urine, but the zoonotic potential of infected cats is still unclear. The objective of this study was to demonstrate if outdoor cats in Germany shed DNA from pathogenic Leptospira species in their urine. As a second aim, antibody prevalence was determined. Methods Two hundred and fifteen outdoor cats were prospectively recruited. Urine samples were tested by real-time PCR targeting the lipL32 gene of pathogenic Leptospira species. Antibody titres against eight serovars (Australis, Autumnalis, Bratislava, Canicola, Copenhageni, Grippotyphosa, Pomona, Saxkoebing) belonging to seven serogroups (Australis, Autumnalis, Canicola, Grippotyphosa, Icterohaemorrhagiae, Pomona, Sejroe) were determined by microscopic agglutination test. Results Urine samples from 7/215 cats (3.3%; 95% confidence interval [CI] 0.9-5.7) were PCR-positive. Specific antibodies were detected in 35/195 cats (17.9%; 95% CI: 12.5-23.3) with titres ranging from 1:100 to 1:6400. Australis, Bratislava and Grippotyphosa were the most common serovars. Conclusions and relevance Outdoor cats in Germany can shed DNA from pathogenic Leptospira species. Therefore, outdoor cats should be considered as a possible source of infection for dogs or humans. Further studies are needed to determine the role of Leptospira species as a cause of disease in cats.