Leptospiral Vaccine Research Articles

Protective immunity of recombinant LipL21 and I-LipL21 against Leptospira interrogans serovar Autumnalis N2 infection.

Leptospirosis is a zoonotic disease caused by the spirochete of genus Leptospira with widespread distribution in tropical, subtropical and temperate zones. Leptospirosis is often confused with other febrile illnesses including jaundice, dengue, and malaria. Generally, the disease is often underdiagnosed or misdiagnosed. Though leptospirosis is curable with antibiotic treatment, the laboratory diagnosis of the disease is specialized and open to interpretation with multiple kits available to detect the different serological markers of Leptospira. Moreover, when leptospirosis is misdiagnosed, the disease can lead to multi-organ failure and may have fatal effects. There is a need for strategies to develop vaccines and prevent leptospirosis. In the present study, the immunogenic potential of leptospiral recombinant protein LipL21 (rLipL21) and its truncated form I-LipL21 (rI-LipL21) was evaluated. The recombinant proteins were established in cyclophosphamide treated BALB/c mice model infected with L. interrogans serovar Autumnalis strain N2. The vaccination study showed 66% and 83% survivability among mice immunized with rLipL21 and rI-LipL21 respectively and post-challenge with leptospiral strain N2 compared to control groups that showed 100% lethality. Additionally, a significant increase in antibody levels and cytokine levels (TNF-a, IFN-γ and IL-10) was observed evidencing a marked stimulation of both humoral and cell-mediated immune response in mice immunized with rLipL21/rI-LipL21 compared to whole cell leptospiral lysate (WCL). This study evidenced protective immunization against leptospirosis with rLipL21 and rI-LipL21 recombinant proteins and are potential candidates for the development of leptospiral vaccine.

Advances & challenges in leptospiral vaccine development.

Considerable progress has been made in the field of leptospiral vaccines development since its first use as a killed vaccine in guinea pigs. Despite the fact that the immunity conferred is restricted to serovars with closely related lipopolysaccharide antigen, certain vaccines have remained useful, especially in endemic regions, for the protection of high-risk individuals. Other conventional vaccines such as the live-attenuated vaccine and lipopolysaccharide (LPS) vaccine have not gained popularity due to the reactive response that follows their administration and the lack of understanding of the pathogenesis of leptospirosis. With the recent breakthrough and availability of complete genome sequences of Leptospira, development of novel vaccine including recombinant protein vaccine using reverse vaccinology approaches has yielded encouraging results. However, factors hindering the development of effective leptospiral vaccines include variation in serovar distribution from region to region, establishment of renal carrier status following vaccination and determination of the dose and endpoint titres acceptable as definitive indicators of protective immunity. In this review, advancements and progress made in LPS-based vaccines, killed- and live-attenuated vaccines, recombinant peptide vaccines and DNA vaccines against leptospirosis are highlighted.

Efficacy of leptospiral commercial vaccines on the protection against an autochtonous strain recovered in Brazil.

In swine and bovines, leptospirosis prevention and control is carried out via vaccination of susceptible animals using bacterins. However, the efficiency of leptospirosis vaccines has been questioned. This work aimed to investigate the potency of five leptospirosis vaccines sold commercially in Brazil, challenging the animals with one autochthonous strain of Leptospira, Canicola serovar, denoted LO4, isolated from swine. The standard protocol was followed, and renal carriers of Leptospira were identified among the surviving animals by culture and PCR. Of the five vaccines tested, only two proved effective. None of the surviving animals was positive by culture; however, one animal was positive by PCR. Three of the five vaccines sold commercially in Brazil for the immunization of swine or bovines failed the test of the efficacy to protect the vaccinated animals following challenge with an autochthonous Leptospira strain, Canicola serovar. The two vaccines provided protection against the renal carrier state in the surviving animals. The criteria used to produce leptospirosis bacterins sold commercially in Brazil must be reviewed. The industry should support researches on leptospiral vaccinology to improve the quality of the present vaccines and discover new immunogenic strains, because it is known that vaccination is one of the most important tools to increase the reproduction rates in livestock.

Humoral Response in Naturally Exposed Horses After Leptospiral Vaccination

Leptospirosis causes substantial economic losses in horses. Nevertheless, humoral response postvaccine is not well described and there is a paucity of knowledge regarding the dynamics of the response in naturally infected horses under field conditions. Fifty-four randomly selected mares, 36 vaccinated (group I) and 18 nonvaccinated (II—control), were used. Mares in group I were given two doses of a multivalent (12 antigens) commercial vaccine (D0 and booster on D40). Blood samples were collected (for microscopic agglutination test and enzyme-linked immunosorbent assay) at D0, D7, D14, D21, D30, D60, and D90. Vaccination significantly interfered with the production of antileptospiral antibodies. Seroreactivity increased rapidly, but was of short duration (up to D60). Responses varied among serogroups present in the vaccine, with the most intense responses against those serogroups that were previously circulating in the herd (Icterohaemorrhagiae and Australis). Therefore, we concluded that vaccines elicited a significant humoral response in horses.

Clinical, serological and echocardiographic examination of healthy field dogs before and after vaccination with a commercial tetravalent leptospirosis vaccine

BackgroundLeptospirosis is a re-emerging bacterial zoonosis caused by spirochetes of the genus Leptospira. Severe disease has been reported in dogs in Europe despite vaccination with bivalent Leptospira vaccines. Recently, a tetravalent canine Leptospira vaccine (Nobivac® L4) was licenced in Europe. The goal of this study was to investigate clinical signs, microscopic agglutination test (MAT) titres, haematology, blood biochemistry, cardiac (c) Troponin I levels and echocardiography before and after vaccination with this tetravalent vaccine. Forty-eight healthy dogs were prospectively enrolled and vaccinated twice, 3–4 weeks apart (T0 and T1). Before vaccination (T0) and 16–31 days after the second vaccination (T2), MAT (n = 48), haematology (n = 48), blood biochemistry (n = 36) and cTroponin I measurements (n = 29) were performed, and MAT was repeated 347–413 days after the second vaccination (T3, n = 44). Echocardiography was performed before the first and second vaccination (T0 and T1, n = 24).ResultsMild and transient clinical signs within 5 days following the first and second vaccination occurred in 23% and 10% of the dogs, respectively. Before the first vaccination (T0), all dogs showed negative MAT titres for the tested serovars except for Canicola (50% with titres 100–400). At T2, positive MAT titres to the serovars Canicola (100%), Australis (89%), Grippotyphosa (86%), Bratislava (60%), Autumnalis (58%), Copenhageni (42%), Pomona (12%), Pyrogenes (8%) and Icterohaemorrhagiae (2%) were found. Median to high titres (≥ 400) were most common to the serovar Canicola (92%) and less common to the serovars Australis (41%), Grippotyphosa (21%), Bratislava (12%), Autumnalis (4%), Pyrogenes (4%) and Pomona (2%). At T3, positive MAT titres (titre range: 100–400) were found in 2–18% of the dogs to serovars of the vaccine serogroups and in 2–18% of the dogs to the non-vaccine serovars Pomona, Autumnalis, Pyrogenes and Ballum. Haematology, blood biochemistry, cTroponin I levels and echocardiography results did not change significantly following vaccination.ConclusionsClinical signs following vaccination with Nobivac® L4 were transient and mild in all cases. Seroconversion differed considerably among individual dogs and among the vaccine serogroups.

Discovery of Novel Leptospirosis Vaccine Candidates Using Reverse and Structural Vaccinology.

Leptospira spp. are diderm (two membranes) bacteria that infect mammals causing leptospirosis, a public health problem with global implications. Thousands of people die every year due to leptospirosis, especially in developing countries with tropical climates. Prophylaxis is difficult due to multiple factors, including the large number of asymptomatic hosts that transmit the bacteria, poor sanitation, increasing numbers of slum dwellers, and the lack of an effective vaccine. Several leptospiral recombinant antigens were evaluated as a replacement for the inactivated (bacterin) vaccine; however, success has been limited. A prospective vaccine candidate is likely to be a surface-related protein that can stimulate the host immune response to clear leptospires from blood and organs. In this study, a comprehensive bioinformatics approach based on reverse and structural vaccinology was applied toward the discovery of novel leptospiral vaccine candidates. The Leptospira interrogans serovar Copenhageni strain L1-130 genome was mined in silico for the enhanced identification of conserved β-barrel (βb) transmembrane proteins and outer membrane (OM) lipoproteins. Orthologs of the prospective vaccine candidates were screened in the genomes of 20 additional Leptospira spp. Three-dimensional structural models, with a high degree of confidence, were created for each of the surface-exposed proteins. Major histocompatibility complex II (MHC-II) epitopes were identified, and their locations were mapped on the structural models. A total of 18 βb transmembrane proteins and 8 OM lipoproteins were identified. These proteins were conserved among the pathogenic Leptospira spp. and were predicted to have epitopes for several variants of MHC-II receptors. A structural and functional analysis of the sequence of these surface proteins demonstrated that most βb transmembrane proteins seem to be TonB-dependent receptors associated with transportation. Other proteins identified included, e.g., TolC efflux pump proteins, a BamA-like OM component of the βb transmembrane protein assembly machinery, and the LptD-like LPS assembly protein. The structural mapping of the immunodominant epitopes identified the location of conserved, surface-exposed, immunogenic regions for each vaccine candidate. The proteins identified in this study are currently being evaluated for experimental evidence for their involvement in virulence, disease pathogenesis, and physiology, in addition to vaccine development.

Evaluation of the Leptospira species microscopic agglutination test in experimentally vaccinated cats and Leptospira species seropositivity in aged azotemic client-owned cats.

The objectives of this study were to validate the microscopic agglutination test (MAT) using feline sera, determine cross-reactivity of Borrelia burgdorferi antibodies in the MAT, and evaluate if there is an association between Leptospira species seropositivity in aged (⩾10 years) client-owned cats with and without azotemia (creatinine >2 g/dl). A four-serovar canine leptospiral vaccine was administered to two specific pathogen-free (SPF) cats on days 0 and 14. The MAT was performed intermittently until day 42 for the serovars Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona and Bratislava, with a cut-off value of ⩾1:100. Five purpose-bred cats were infested with wild-caught Ixodes scapularis adults with an average B burgdorferi infection rate of 50%, and tested for antibodies against B burgdorferi C6 peptide and DNA in skin biopsies, as well as by MAT. Sera from 66 azotemic and 75 non-azotemic cats ⩾10 years of age were tested for Leptospira species antibodies using the MAT and results were compared by the χ(2) test. Both SPF cats seroconverted by week 3 and formed antibodies against at least one serovar. There was no cross-reactivity in the MAT using samples from cats with antibodies to B burgdorferi. MAT results were positive for 4/66 azotemic cats and 8/75 non-azotemic cats; these results were not statistically different. The MAT can be interpreted using feline serum and does not appear to cross-react in cats with B burgdorferi antibodies. There was no association between Leptospira species MAT results and azotemia in this group of aged client-owned cats but further studies are needed to determine if leptospirosis contributes to feline chronic kidney disease.

Effectiveness of a commercial leptospiral vaccine on urinary shedding in naturally exposed sheep in New Zealand

L. borgpetersenii serovar Hardjo and L. interrogans serovar Pomona are endemic in New Zealand sheep. An effective vaccine and vaccination strategy would protect both humans and livestock.Four to 12 lambs were selected from each of eight farms (total=84, vaccinated group), while four to 16 lambs (total=98) served as unvaccinated controls. A commercial Hardjo/Pomona vaccine was given at 1–6 weeks of age, 5–11 weeks later and 33–67 weeks later on seven farms and at 18 weeks of age and 5 weeks later on the eighth farm. Vaccinates and controls were grazed together. Blood was regularly collected from the control group to assess flock exposure. Urine was collected from both groups 26–82 weeks after the second vaccination and tested by quantitative PCR.Seroprevalence in controls at the time of urine sampling ranged from 2.7 to 98.2% for Hardjo and from 0 to 54.1% for Pomona with seroconversion occurring 13 to 67 weeks after the second vaccination in all but one farm where exposure had happened by the time of vaccination. The shedding prevalence adjusted for clustering in farms was 45.1% [95% CI 17.6–72.7] (for an observed number of 50/98) in the control animals and 1.8% [95% CI 0.0–10.1] (for an observed number of 5/84) in the vaccinated animals. The vaccine was 100% effective on five farms where animals were vaccinated before 12 weeks of age and before natural exposure occurred, but the effectiveness was 80% [0–97] on one farm where the lambs were exposed before vaccination and 65% [9–87] to 80% [0–97] on one farm where the animals were fully vaccinated by 24 weeks of age. The overall vaccine effectiveness was 86.3% [63.6–94.8%] despite maternal antibodies in some flocks at first vaccination. Vaccination timing seemed to be crucial in achieving optimum reduction in shedding in urine in vaccinated sheep.

Improved Alum Containing Adjuvant by Vitamin A for Enhancing Immune Responses and Efficacy of Leptospira Vaccine in Hamster Model

Use of suitable adjuvant is one of the priorities in vaccine and immune modulation, stimulation and potentiating. Vaccination is effective for prevention and treatment of bacterial diseases including Leptospirosis. In the present study, we prepared Leptospiral vaccine with Alum, modified Alum adjuvant (mAlum) and without adjuvant. Vaccination was done; then we evaluated the immune responses by isolating the splenocytes and sera for interleukin (IL) profiles (the highest level of all cytokines except IL-4 and IL-12 was obtained in the mAlum antigen group at week 7 post-treatment). Moreover, the expression of all evaluated cytokines in the mAlum group was greater than in the other groups at week 62. Significant increases in antibody titters were noted in the mAlum and Alum group, challenged interaperitionelly with a lethal dose of virulent and monitored pathological lesion that moderate to severe lesions with score 3 were observed in the control group while the animals immunized with mAlum-antigen and Alum-antigen displayed slight to mild lesions with an average score of 0.5. The results demonstrated that modified Alum Adjuvants are better than Alum adjuvants as revealed by the enhanced long-term antibody responses. None of the vaccinated animals died from the challenge experiment 84 day post-injection except those vaccinated with saline vaccine. We assume that these observations affirmatively assign a pivotal vitamin a role to adjuvant formulation and preparation in type and extent of immune responses raised in the vaccinated animals.

A novel octavalent combined Erysipelas, Parvo and Leptospira vaccine provides (cross) protection against infection following challenge of pigs with 9 different Leptospira interrogans serovars.

BackgroundIn the present study the efficacy and cross protection of a novel ready-to-use combination vaccine, Porcilis® Ery + Parvo + Lepto, against swine leptospirosis, was investigated. The octavalent vaccine contains inactivated antigens of Erysipelothrix rhusiopathiae, Parvovirus and 6 Leptospira interrogans sensu lato strains of serogroups Canicola, Icterohaemorrhagiae, Grippotyphosa, Australis (Bratislava), Pomona and Tarassovi. In this study ninety pigs were vaccinated twice with Porcilis® Ery + Parvo + Lepto at six and ten weeks of age and ninety age and source-matched animals served as unvaccinated control. Two weeks after booster vaccination, groups of vaccinated and control pigs (ten pigs per group) were challenged with fresh virulent cultures of either of the nine different challenge strains. Compared to the vaccine strains the challenge strains were heterologous strains of the same serovar or of a different serovar within the same serogroup. The challenge strains tested were of serovar Canicola, Icterohaemorrhagiae, Copenhageni (serogroup Icterohaemorrhagiae), Grippotyphosa, Bananal/Liangguan (serogroup Grippotyphosa), Pomona, Tarassovi and Vughia (serogroup Tarassovi).ResultsAfter the different challenges most control animals became leptospiraemic for 2–7 days. The vaccinated pigs remained blood culture negative except for two animals after serovar Icterohaemorrhagiae and two animals after serovar Tarassovi challenge which became leptospiraemic for only 1 day. The incidence of Leptospiraemia (as determined by blood culture) was significantly less in vaccinates compared to the controls after all challenges. The vaccine also prevented renal infection and urinary shedding after serovar Canicola challenge. The other serovars did not induce detectable renal infection or urinary shedding.ConclusionThe present study demonstrates that the new combination vaccine Porcilis® Ery + Parvo + Lepto induces significant (cross) protection against nine different serovars within the serogroups Canicola, Icterohaemorrhagiae, Grippotyphosa, Australis (Bratislava), Pomona and Tarassovi.

Incidence rates and risk factors for owner-reported adverse events following vaccination of dogs that did or did not receive a Leptospira vaccine.

To determine incidence rates (IRs) and potential risk factors for owner-reported adverse events (AEs) following vaccination of dogs that did or did not receive a Leptospira vaccine. Observational, retrospective cohort study. 130,557 dogs. Electronic records of mobile veterinary clinics from June 2012 to March 2013 were searched to identify dogs that received ≥ 1 vaccine in a given visit. Signalment data, vaccinations received, medications administered, and owner-reported clinical signs consistent with AEs that developed ≤ 5 days after vaccination were recorded. Associations between potential risk factors and owner-reported AEs were evaluated by logistic regression analysis. The IR/10,000 dogs for owner-reported postvaccination AEs was 26.3 (95% CI, 23.6 to 29.2), whereas that for dogs that received a Leptospira vaccine alone or with other vaccines was 53.0 (95% CI, 42.8 to 64.9). Significant factors for increasing or decreasing risk of AEs were as follows: receiving a Leptospira vaccine (adjusted OR, 2.13), age at vaccination 1 to < 7 or ≥ 7 years (vs a referent of < 6 months; adjusted OR, 0.54 and 0.44, respectively), and weight 13.6 to < 22.7 kg (30 to < 50 lb) and 22.7 to < 45.5 kg (50 to 100 lb [vs a referent of < 4.5 kg {10 lb}]; adjusted OR, 0.48 and 0.55, respectively). Hypersensitivity reactions were rare (IR, 6.5/10,000 dogs), and IRs for these events did not differ significantly between dogs vaccinated with or without a Leptospira component. The overall IR for owner-reported postvaccination AEs was low. Results suggested vaccination against Leptospira (an organism that can cause fatal disease) is safe in the majority of cases, slightly increasing the risk of owner-reported AEs but not associated with a significant increase in hypersensitivity reactions, compared with other vaccinations administered.

Safety and efficacy of a new octavalent combined Erysipelas, Parvo and Leptospira vaccine in gilts against Leptospira interrogans serovar Pomona associated disease and foetal death

The safety and protective efficacy of a new octavalent combination vaccine containing inactivated Erysipelothrix rhusiopathiae, Parvovirus, and Leptospira interrogans (sensu lato) serogroups Canicola, Icterohaemorrhagiae, Australis (Bratislava), Grippotyphosa, Pomona and Tarassovi – Porcilis® Ery+Parvo+Lepto – was evaluated in laboratory studies and under field conditions.The safety (2× overdose and repeated dose) was tested in 26 gilts. In this study, neither vaccine related temperature increase nor other systemic reactions were observed after intramuscular vaccination. No local reactions were observed except for one animal that had a small local reaction (2cm diameter) that lasted for 5 days after the third vaccination.Efficacy was tested in 40 gilts. A group of 20 gilts was vaccinated at 20 and 24 weeks of age with Porcilis® Ery+Parvo+Lepto and a group of 20 age- and source-matched animals served as the control group. The gilts were inseminated at 41 weeks or 66 weeks of age and were challenged with serovar Pomona 10 weeks after insemination, corresponding to 6 months (n=2×10) and 12 months (n=2×10) after the last vaccination. After both the 6- and 12-month challenges the control animals developed clinical signs (fever, lethargy and anorexia) and leptospiraemia as determined by positive blood culture. In addition, both the 6- and 12-month challenges resulted in death of 21% and 27% of the total number of foetuses in the control groups, respectively. Clinical signs and leptospiraemia were statistically significantly lower in vaccinated gilts after both the 6- and 12-month challenges. In addition, foetal death was statistically significantly lower (3% and 2%, respectively) in vaccinated gilts after both the 6- and 12 month challenges.The vaccine was tested further under field conditions on a Portuguese farm with a history of an increasing abortion rate associated with a Leptospira serovar Pomona infection (confirmed by PCR and serology). This study was designed as an observational-longitudinal field study. At the start of the study, all breeding sows and replacement gilts on the farm were vaccinated twice with Porcilis® Ery+Parvo+Lepto at an interval of 4 weeks. Starting six months after the primary vaccination schedule, the animals were re-vaccinated during the second week of every subsequent lactation. New replacement gilts were vaccinated using the same schedule. After vaccination, the abortion rate reduced rapidly from 12.6% in winter months of 2012 (December 2011 to March 2012) to 0.5% in winter months of 2013, a statistical significant decrease of 96%. The total number of abortions on the farm decreased from 55 in 2012 to 6 in 2013. Thereafter, the abortion rate remained stable and in the period December 2013 to April 2014 was still low (0.6%).In conclusion, the present studies demonstrate that the octavalent Porcilis® Ery+Parvo+Lepto vaccine can be safely used in gilts and sows and induces significant protection, for the duration of at least one year, against serovar Pomona induced clinical signs, leptospiraemia and foetal death. Protection against Pomona associated reproductive failure was confirmed under field conditions where a significant reduction in abortion rate was observed.

Genetic stability of vaccine strains by multilocus sequence typing and pulsed-field gel electrophoresis analysis: Implications for quality control of the leptospiral vaccine

Quality control of vaccine strains is directly associated with the safety and efficacy of inactivated whole bacterial vaccines. The assessment of genetic stability is one of the essential elements to guarantee the quality of vaccine strains. The multiple-valence inactivated leptospiral vaccine, comprising the main circulating serogroups, has played an important role in the control of Leptospira infection in China. In the present study, to assess the genetic stability of vaccine strains and develop novel quality control tests that enhance and extend the existing procedures, 7 Chinese leptospiral vaccine strains were characterized during in vivo and in vitro passages by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) analysis. The seven vaccine strains were found to have distinct sequence types (STs) and PFGE profiles. Further analysis showed that the ST and PFGE pattern of each vaccine strain, after in vivo or serial in vitro passages (up to 20 passages), were identical to those of the initial strain, demonstrating that these strains were genetically stable and homogeneous. Taken together, PFGE and MLST provide a reproducible and reliable means for confirming the identity and genetic stability of vaccine seeds, suggesting that these approaches can be used to evaluate the quality of leptospiral vaccine strains.

Molecular identification of the ompL1 gene within Leptospira interrogans standard serovars.

Leptospirosis, caused by infection with pathogenic Leptospira species, is one of the most prevalent zoonotic diseases in the world. Current leptospiral vaccines are mainly multivalent dead whole-cell mixtures made of several local dominant serovars. Therefore, design and construction of an efficient recombinant vaccine for leptospirosis control is very important. OmpL1 is an immunogenic porin protein that could be of special significance in vaccination and serodiagnosis for leptospirosis. Three strains belonging to pathogenic L. interrogans were analyzed. The specific primers for proliferation of the ompL1 gene were designed. The amplified gene was cloned. In order to investigate the ompL1 nucleotide sequence and homological analysis of this gene, ompL1 genes cloned from standard vaccinal Leptospira serovars prevalent in Iran were sequenced and cloned. PCR amplification of the ompL1 gene using the designed primers resulted in a 963 bp ompL1 gene product. The PCR based on the ompL1 gene detected all pathogenic reference serovars of Leptospira spp. tested. Based on alignment and phylogenetic analysis, although the ompL1 nucleotide sequence was slightly different within three vaccinal serovars (100%-85% identity), amino acid alignment of the OmpL1 proteins revealed that there would be inconsiderable difference among them. The ompL1 gene of the three isolates was well conserved, differing only by a total of 6 bp and the proteins by 2 amino acids. The cloned gene could be further used for expression and recombinant OmpL1 as an efficient and conserved antigen, and may be a useful vaccine candidate against leptospirosis in our region.

Comparision of efficacy of two experimental bovine leptospira vaccines under laboratory and field

Two different inactivated vaccines for bovine leptospirosis were prepared with five different Leptospira serovars namely australis, ballum, hardjo, hebdomadis and pomona and adjuvanted with Montanide ISA 206 (Vaccine-I) or adjuvanted with Aluminium hydroxide gel (Vaccine-II) to evaluate the efficacy of the vaccines. The immunogenicity of the vaccines was studied in rabbits under experimental condition and in cattle under field condition for a period of 180 days. Antibody response against five different leptospira serovars ranged from 6.84log2 to 9.64log2 (GM-MAT titres) in rabbits at 180 days after vaccination with vaccine I, whereas in vaccine II, the titre value ranged from 5.64log2 to 7.44log2. In the case of cattle under field condition, vaccine I showed GM-MAT titres of 6.84log2 to 7.69log2 against five different serovars at 150 days following vaccination. Such titres were maintained following vaccination with vaccine II for 120 days only. It is concluded that vaccine I is a better vaccine than vaccine II. However both the vaccines showed high immune response of 5.64log2 at six months of immunization. Vaccination did not cause any adverse reaction in the vaccinated cattle.

Vaccine‐Associated Leptospira Antibodies in Client‐Owned Dogs

BackgroundLong‐term microscopic agglutination test (MAT) results after vaccination with 4‐serovar Leptospira vaccines are not available for all vaccines used in client‐owned dogs.Hypothesis/ObjectivesTo determine antibody responses of client‐owned dogs given 1 of 4 commercially available Leptospira vaccines.AnimalsHealthy client‐owned dogs (n = 32) with no history of Leptospira vaccination for at least the previous year.MethodsDogs were given 1 of 4 Leptospira vaccines on week 0 and then approximately on week 3 and week 52. Sera were collected before vaccine administration on week 0 and then within 3 days of week 3, within 2 days of week 4, and approximately on weeks 7, 15, 29, 52, and 56. Antibody titers against Leptospira serovars bratislava, canicola, grippotyphosa, hardjo, icterohemorrhagiae, and pomona and were determined by MAT.ResultsWhen compared among vaccines, MAT results varied in maximal titers, the serovars inducing maximal titers, and the time required to reach maximal titers. Each vaccine induced at least some MAT titers ≥1 : 800. Most dogs were negative for antibodies against all serovars 1 year after vaccination, and anamnestic responses were variable.Conclusions and Clinical ImportanceDogs vaccinated with Leptospira vaccines have variable MAT titers over time, and antibodies should not be used to predict resistance to Leptospira infection. MAT titers ≥1 : 800 can develop after Leptospira spp. vaccination, which can complicate the clinical diagnosis of leptospirosis.

Leptospira cases and vaccination habits within UK vet-visiting dogs

Veterinary RecordVolume 174, Issue 11 p. 278-278 Research Leptospira cases and vaccination habits within UK vet-visiting dogs C. Ball BSc., Hons, Corresponding Author C. Ball BSc., Hons [email protected] Department...

Signalment Changes in Canine Leptospirosis between 1970 and 2009

BackgroundPrevious studies have identified large breed, male, outdoor dogs of herding or working groups to be at increased risk for Leptospira infection. Exposure risk factors may change over time, altering the signalment of dogs most commonly diagnosed with leptospirosis.ObjectivesThe objectives of this study were to evaluate possible signalment changes by decade in canine leptospirosis cases diagnosed at university veterinary hospitals in the United States and Canada using reports to the Veterinary Medical DataBase (VMDB) over a 40‐year period (1970–2009).AnimalsOne thousand and ninety‐one dogs with leptospirosis diagnosed among 1,659,146 hospital visits.MethodsHospital prevalence of leptospirosis by decade was determined by age, sex, weight, and breed groups. Multivariable logistic regression models were created to evaluate the association between variables and the odds of disease for each decade.ResultsVeterinary Medical DataBase hospital prevalence of leptospirosis in dogs, after a marked decrease in the 1970s and low rates in the 1980s, began increasing in the 1990s. Hospital prevalence significantly increased in dogs between 2 and 9.9 years of age (P < .05) and in male dogs (P < .05) in each decade since the 1980s. Among weight groups in the most recent decade (2000–2009), dogs weighing <15 pounds had the greatest odds of being diagnosed with leptospirosis (P = .003).Conclusions and Clinical ImportanceHospital prevalence rates by age, weight, sex, and breed groups differed by decade. These changes may reflect changes in exposure risk, Leptospira vaccination practices for dogs, or both.

Development and validation of a serological potency test for the release of Leptospira vaccines – Requirements in the European Union

Both European Pharmacopoeia Monograph 01/2008:0447 “Canine Leptospirosis vaccine (inactivated)” and the more recent Monograph 01/2008:1939 “Bovine Leptospirosis vaccine (inactivated)” explicitly allow for a sero-response test to assess batch potency. Test setup and requirements for in vivo and in vitro validation are described. Furthermore, the two main strategies to assess batch potency and their specific demands are addressed.

Opportunities and strategies to further reduce animal use for Leptospira vaccine potency testing

Hamsters are routinely infected with virulent Leptospira for two purposes in the regulation of biologics: the performance of Codified potency tests and maintenance of challenge culture for the Codified potency tests. Options for reducing animal use in these processes were explored in a plenary lecture at the “International Workshop on Alternative Methods for Leptospira Vaccine Potency Testing: State of the Science and the Way Forward” held at the Center for Veterinary Biologics in September 2012. The use of validated in vitro potency assays such as those developed by the U.S. Department of Agriculture for Leptospira (L.) canicola, Leptospira grippotyphosa, Leptospira pomona, and Leptospira icterohaemorrhagiae rather than the Codified hamster vaccination–challenge assay was encouraged. Alternatives such as reduced animal numbers in the hamster vaccination–challenge testing were considered for problematic situations. Specifically, the merits of sharing challenge controls, reducing group sizes, and eliminating animals for concurrent challenge dose titration were assessed. Options for maintaining virulent, stable cultures without serial passage through hamsters or with decreased hamster use were also discussed. The maintenance of virulent Leptospira without the use of live animals is especially difficult since a reliable means to maintain virulence after multiple in vitro passages has not yet been identified.