Efficacy of an inactivated recombinant vaccine encoding a fimbrial protein of Pasteurella multocida B:2 against hemorrhagic septicemia in goats

This study was carried out to determine the antibody responses and protective capacity of an inactivated recombinant vaccine expressing the fimbrial protein of Pasteurella multocida B:2 following intranasal vaccination against hemorrhagic septicemia in goats. Goats were vaccinated intranasal with 10⁶ CFU/mL of the recombinant vaccine (vaccinated group) and 10⁶ CFU/mL of pET32/LIC vector without fimbrial protein (control group). All three groups were kept separated before all goats in the three groups were challenged with 10⁹ CFU/mL of live pathogenic P. multocida B:2. During the course of study, both serum and lung lavage fluid were collected to evaluate the antibody levels via enzyme-linked immunosorbent assay. It was found that goats immunized with the inactivated recombinant vaccine developed a strong and significantly (p < 0.05) higher specific IgA and IgG responses in both serum and lung lavage fluid samples compared to the control and unvaccinated groups. Following intratracheal challenge, the rate of isolation was 17% for the vaccinated group, 67% for the control group and 100% for the unvaccinated group. However, none of the goat from the vaccinated group had P. multocida B:2 in the liver, tonsil and heart. Therefore, the study revealed that an inactivated recombinant vaccine significantly provides significant protection against high dose challenge and enhances the stimulation of the local and systemic immunities.     

Development of a novel LAMP diagnostic method for visible detection of swine <Emphasis Type="Italic">Pasteurella multocida</Emphasis>

A set of four specific primers for six regions of kmt1 gene from a species specific region was designed for developing the loop-mediated isothermal amplification diagnostic method of swine Pasteurella multocida (Pm-LAMP). After the Pm-LAMP was carried out at 63°C for 1 h, the LAMP products could be visually confirmed using fluorescent dyes as detection reagent under UV-illumination. In sensitivity, the detection limit of the Pm-LAMP was 10 cfu/mL, and was 1 log less than that of the PCR method. In specificity, the Pm-LAMP did not amplify genomic DNA of swine common respiratory pathogens. Furthermore, based on results for clinical swab samples (n = 31) using PCR detection as golden standard, relative sensitivity of the Pm-LAMP was 100%, relative specificity of the Pm-LAMP was 90.9%, and percentage of observation agreement was 93.5% (Kappa = 0.85). The Pm-LAMP method should be a useful diagnostic tool for rapid and visible detection of swine Pasteurella multocida.     

Proliferation and transmission patterns of <Emphasis Type="Italic">Pasteurella multocida</Emphasis> B:2 in goats

This report describes the proliferation and transmission patterns of Pasteurella multocida B:2 among stressful goats, created through dexamethasone injections. Thirty seven clinically healthy adult goats were divided into three groups consisted of 15 goats in group A, 11 goats in group B and the remaining 11 in group C. At the start of the study, all goats of group A were exposed intranasally to 1.97 × 10¹⁰ CFU/ml of live P. multocida B:2. Dexamethasone was immediately administered intramuscularly for 3 consecutive days at a dosage rate of 1 mg/kg. The exposed goats were observed for signs of HS for a period of 1 month. At the end of the 1-month period, 11 goats from group B were introduced into and commingled with the surviving goats of group A before all goats from both groups were immediately injected intramuscularly with dexamethasone for 3 consecutive days. The treatment with dexamethasone was then carried out at monthly interval throughout the 3-month study period. Goats of group C were kept separately as negative control. Three surviving goats from each group were killed at 2-week interval for a complete post-mortem examination. Two (13%) goats of group A were killed within 24 hours after intranasal exposure to P. multocida B:2 while another two (13%) goats from the same group were killed on day 40, approximately 10 days after the second dexamethasone injection. All four goats showed signs and lesions typical of haemorrhagic septicaemia. Bacteraemia was detected in 3 goats of group A that were having rectal temperature higher than 41°C. The P. multocida B:2 isolation pattern was closely associated with dexamethasone injections when significantly (p < 0.05) higher rate of isolations from both groups were observed after each dexamethasone injection. Transmission of P. multocida B:2 from goats of group A to group B was successful when P. multocida B:2 was isolated from goats of group B for a period of 28 days. There was a strong correlation between dexamethasone injections, rate of bacterial isolation and serum cortisol level. The IgG level showed an increasing trend 2 weeks after exposure to P. multocida B:2 and remained high throughout the study period.     

Mouse model of haemorrhagic septicaemia: dissemination and multiplication of Pasteurella multocida B:2 in vital organs after intranasal and subcutaneous challenge in mice

Haemorrhagic septicaemia (HS) is an endemic disease of bovines, occuring in most tropical regions of Asia and Africa. In the present study, the suitability of using mice to study pathogenesis of HS was assessed using mortality, mean death time and bacterial multiplication in vital organs after infection with live P multocida. Mice were infected with 10⁵, 10³ and 10¹?cfu of P. multocida B:2 via intranasal and subcutaneous routes along with control groups. Bacterial multiplication in lung, liver and spleen of mice were determined at 24 h interval after intranasal and subcutaneous challenge. More than 80 % of challenged mice died within 48 h of inoculation, irrespective of the dose and route of inoculation. A heavy bacterial load (up to 10⁸?cfu) was observed in lung, liver and spleen of mice titrated at 24 h and following death of mice. Results of the present study indicate that even ten bacteria are enough to cause mortality in mice and the organism multiplies rapidly in respiratory epithelium and disseminated to other vital organs viz liver and spleen suggesting the important role of mouse model in investigating the pathogenesis and challenge studies during vaccine development.     

Comparison of the pathogensis of two isolates of <Emphasis Type="Italic">Besnoitia caprae</Emphasis> in inbred BALB/c mice

This study was performed to evaluate the infectivity of bradyzoites of two Besnoitia caprae isolates, BC-1 and BC-2, to inbred BALB/c mice. Each group of inbred BALB/c mice was inoculated intraperitoneally with 1 × 10³, 1 × 10⁴, 1 × 10⁵, 5 × 10⁵ and 1 × 10⁶ of one of the two isolates of B. caprae bradyzoites. The mice were monitored daily for a period of 40 days for survival. After death of each mice, several passages from its peritoneal washing and tissues were analyzed using ribosomal DNA-specific PCR assay. Marked differences in pathogenicity between the isolates were seen. All the inbred BALB/c mice infected with BC-2 survived but all the mice that were administered with 1 × 0⁵, 5 × 10⁵ and 1 × 10⁶ BC-1 bradyzoites were died within 4–9 days post-infection (DPI). Histopathological examination of the tissues of the dead mice revealed hyperemia and necrosis with presence of mononuclear and polymorphonuclear cell infiltration in myocardium, spleen and intestines together with interstitial pneumonia and peritonitis. All inbred BALB/c mice in the 1 × 10³ and 1 × 10⁴ groups of BC-1 inoculated mice survived and they were euthanized after 40 DPI. Chronic inflammation with infiltration of mononuclear cells was evident in myocardium, spleen, alveolar septa of the lungs of most of the examined tissues with hemorrhagic enteritis in the mice infected with 1x10⁶ bradyzoites. The mice infected with different doses of BC-2 were euthanized after 40 DPI and no lesion was seen in histopathological sections of their organs. All peritoneal washings and examined tissues were PCR positive in BC-1 group. This experiment is the first report to show inbred BALB/c mice as a relevant model for B. caprae and demonstrates that this strain of inbred BALB/c mice is a suitable animal model for biological studies and examination of pathogenesis for this species of Besnoitia. The present findings also provide evidence for significant differences between the two isolates of B. caprae.     

Protective effect following intranasal exposure of goats to live Pasteurella multocida B:2

This study aimed to determine the effect of intranasal exposure to low doses of Pasteurella multocida B:2 on survival of goats challenged with high doses of the same organism. Eighteen goats were selected and divided into three groups. Goats of group 1 were exposed intranasally twice, with a two-week interval, to 7× 10⁶ cfu/ml of live P. multocida B:2. Goats of group 2 were not exposed to P. multocida B:2 but were kept together with the exposed group 1. Goats of group 3 remained as unexposed controls and were kept separated from the other two groups. Serum samples were collected at weekly intervals to determine the antibody levels. At week 5 post exposure, all goats were challenged subcutaneously with 3.7× 10¹⁰ cfu/ml of live P. multocida B:2. Following challenge exposure, 8 (67%) goats (4 goats from each of groups 1 and 2) were killed owing to haemorrhagic septicaemia. Four goats were killed peracutely within 48 h post challenge, while the other four goats were killed acutely between 2 and 4 days post challenge. None of the goats of group 3 were killed for haemorrhagic septicaemia. Goats of groups 1 and 2 showed significantly (p<0.05) higher antibody levels following the first intranasal exposure to P. multocida B:2. However, only group 1 retained the significantly (p<0.05) high antibody levels following a second intranasal exposure, and remained significantly (p<0.05) higher than groups 2 and 3 at the time of challenge. P. multocida B:2 was successfully isolated from various organs of goats that were killed between 1 and 4 days post challenge.     

Standardization of the outbred BALB/c mice as a suitable animal model for <Emphasis Type="BoldItalic">Besnoitia caprae</Emphasis> studies

The Apicomplexan parasite Besnoitia caprae infects wild and domestic goats. Knowledge on Besnoitia caprae specially an optimized animal model is sparse. Experimental infections with tachyzoites of BC-Pars obtained from BALB/c mice were conducted in outbred mice to determine the infectivity and LD50 of Besnoitia caprae. Six groups of five mice were intraperitoneally infected with 12.5 × 10³, 25 × 10³, 5 × 10⁴, 1 × 10⁵ and 2 × 10⁵ tachyzoites and a control inoculum of DMEM, respectively. Although morbidity and mortality were observed in all groups, two mice in the 12.5 × 10³ group showed alopecia and skin lesions on 60 days post-infection (DPI). Histopathological and molecular examination of skins confirmed B. caprae infection. The LD₅₀ was calculated as 25 × 10^3.2 tachyzoites per mouse. The results indicate that outbred BALB/c mice can be used as a suitable model of besnoitiosis and to screen candidate treatments and to test the efficacy of vaccines for besnoitiosis.     

Factors influencing production of hygienic raw milk by small scale dairy producers in selected areas of the Jaffna district,Sri Lanka

The objective of the study was to investigate the influence of dairy cow management techniques and milking methods on hygienic quality of raw milk. Total Bacterial Count (TBC) and Total Coliform Colonies (TCC) were studied to determine the effects. Investigations were carried out in fifty dairy farms from August 2007 to December 2007. The mean TBC and TCC for the herds with comparatively good and poor management practices were 0.9 × 10⁵ cfu/ml and 0.2 × 10³/ml and 99 × 10⁵ cfu/ml and >180 × 10³/ml, respectively. The overall mean TBC (22 × 10⁵ cfu/ml) and TCC (47 × 10³/ml) obtained in this study exceeded the internationally recommended levels for TBC (10⁵ cfu/ml) and TCC (<1,000/ml). The overall results obtained suggested that the raw milk tested was of poor hygienic quality with the presence of a great variability among milk samples.     

Preparation and evaluation of chicken embryo-adapted fowl adenovirus serotype 4 vaccine in broiler chickens

The current study was planned to develop an efficient vaccine against hydropericardium syndrome virus (HSV). Currently, formalin-inactivated liver organ vaccines failed to protect the Pakistan broiler industry from this destructive disease of economic importance. A field isolate of the pathogenic hydropericardium syndrome virus was adapted to chicken embryos after four blind passages. The chicken embryo-adapted virus was further serially passaged (12 times) to get complete attenuation. Groups of broiler chickens free from maternal antibodies against HSV at the age of 14 days were immunized either with 16th passage attenuated HSV vaccine or commercially formalized liver organ vaccine. The antibody response, measured by enzyme-linked immunosorbent assay was significantly higher (P < 0.05) in the group immunized with the 16th passage attenuated HSV vaccine compared to the group immunized with liver organ vaccine at 7, 14, and 21 days post-immunization. At 24 days of age, the broiler chickens in each group were challenged with 10^3.83 embryo infectious dose₅₀ of pathogenic HSV and were observed for 7 days post-challenge. Vaccination with the 16th passage attenuated HSV gave 94.73% protection as validated on the basis of clinical signs (5.26%), gross lesions in the liver and heart (5.26%), histopathological lesions in the liver (1.5 ± 0.20), and mortality (5.26%). The birds inoculated with liver organ vaccine showed significantly low (p < 0.05; 55%) protection estimated on the basis of clinical signs (40%), gross lesions in the liver and heart (45%), histopathological lesions in the liver (2.7 ± 0.72), and mortality (35%). Birds in the unvaccinated control group showed high morbidity (84%), mortality (70%), gross (85%), and histopathological lesions (3.79 ± 0.14) with only 10% protection. In conclusion, this newly developed HSV vaccine proved to be immunogenic and has potential for controlling HSV infections in chickens.     

Identification of immunogenic proteins associated with protection against haemorrhagic septicaemia after vaccination of calves with a live-attenuated aroA derivative of Pasteurella multocida B:2

Pasteurella multocida serotype B:2 is the causative agent of haemorrhagic septicaemia (HS), a fatal disease of cattle and buffaloes. As a step towards the identification of individual antigens that may protect against HS, proteins present in a sonicated cell extract (SCE) and outer-membrane protein (OMP) preparation of a wild-type P. multocida serotype B:2 were investigated by immunoblotting with sera from calves which had been protected against challenge with a virulent strain of P. multocida B:2 by vaccination with a live-attenuated aroA derivative of the challenge strain. Five proteins in SCE, of approximately 50, 37, 30, 26 and 16 kDa, were recognised by the sera. In an OMP preparation, two bands, at 37 and 50 kDa, were recognised as strongly immunogenic. Mass spectrometry analysis of proteins corresponding in size to those detected by immunoblotting identified the 37 kDa band as OmpA, but the band at 50 kDa was not identified with certainty. A major 30 kDa OMP, identified as OmpH, was not strongly immunogenic.     

Enhancing mucosal immunity in mice by recombinant adenovirus expressing major epitopes of porcine circovirus-2 capsid protein delivered with cytosine-phosphate-guanosine oligodeoxynucleotides

A recombinant replication-defective adenovirus expressing the major epitopes of porcine circovirus-2 (PCV-2) capsid protein (rAd/Cap/518) was previously constructed and shown to induce mucosal immunity in mice following intranasal delivery. In the present study, immune responses induced by intranasal immunization with a combination of rAd/Cap/518 and cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODN) were evaluated in mice. The levels of PCV-2-specific IgG in serum and IgA in saliva, lung, and intestinal fluids were significantly higher in the group immunized with rAd/Cap/518 and CpG ODN than animals immunized with rAd/Cap/518 alone. The frequencies of IL-2-secreting CD4⁺ T cells and IFN-γ-producing CD8⁺ T cells were significantly higher in the combined immunization group than mice immunized with rAd/Cap/518 alone. The frequencies of CD3⁺, CD3⁺CD4⁺CD8^-, and CD3⁺CD4^-CD8⁺ T cells in the combined immunization group were similar to that treated with CpG ODN alone, but significantly higher than mice that did not receive CpG ODN. PCV-2 load after challenge in the combined immunization group was significantly lower than that in the phosphate-buffered saline placebo group and approximately 7-fold lower in the group treated with CpG ODN alone. These results indicate that rAd/Cap/518 combined with CpG ODN can enhance systemic and local mucosal immunity in mice, and represent a promising synergetic mucosal vaccine against PCV-2.     

Effect of source and dose of probiotics and exogenous fibrolytic enzymes (EFE) on intake,feed efficiency,and growth of male buffalo (<Emphasis Type="BoldItalic">Bubalus bubalis</Emphasis>) calves

Probiotics of Lactobacillus acidophilus, Saccharomyces cerevisiae, and Aspergillus niger and three commercial exogenous fibrolytic enzymes (EFE) were tested in vitro to select best source and optimum dose followed by in vivo studies on male buffalo calves. Bacterial (P < 0.001) and protozoal population were increased significantly (P < 0.001) with probiotics and EFE. In vitro dry matter digestibility was more (P < 0.001) on L. acidophilus and then on S. cerevisiae. Dose required for L. acidophilus and S. cerevisiae probiotics was 1 × 10⁹ and 3 × 10⁹ cfu/flask, respectively. Cellulase and xylanase were effective at 4,000 and 12,500 IU/kg DM. In vitro cell wall digestibility was increased (P < 0.001) when probiotics and EFE were used together. Best source and optimum dose of probiotics and EFE were fed to 18 male buffalo calves with concentrate supplement (CS). Calves were randomly divided into three groups either without probiotics and EFE (CG) or with probiotics (EG₁) or probiotics combined with EFE (EG₂) on wheat straw diet. Organic matter, neutral detergent fiber, and acid detergent fiber digestibility was improved significantly. Average daily weight gain (ADG) and feed efficiency were significantly higher (P < 0.001) in EG₂ than EG₁ or CG. Final body weight was 4% and 12% and feed efficiency was 2.6% or 1.6% more (P < 0.001) in EG₂ compared to CG or EG₁, respectively. Fortification of CS with probiotics and EFE together had more impact on FE and ADG in buffalo calves.     

Passive protection of mice with antiserum to neuraminidase fromPasteurella multocida serotype A:3

Antiserum to a partially purified neuraminidase fromPasteurella multocida, type A:3, was adsorbed with protease-digestedP. multocida type 3 lipopolysaccharide (LPS) to remove LPS immunoreactivity. The LPS-adsorbed antineuraminidase caused a 77% reduction in the neuraminidase activity of homologousP. multocida in anin vitro enzyme neutralization test. All 14 mice passively immunized with the adsorbed antineuraminidase were protected against challenge infection with homologousP. multocida in a mouse protection test. Ten out of 14 mice in one group that received antisera containing antibodies to both neuraminidase and LPS were protected. In contrast, only 1 out of 14 mice that were immunized with pre-immune serum survived the challenge. These results suggest that antiserum toP. multocida neuraminidase was, at least partly, responsible for the protection observed in this study. Neuraminidase may be one of the immunogenic protective proteins present in aqueous extracts ofPasteurella multocida.     

Supplementation of cottonseed,linseed, and noug seed cakes on feed intake,digestibility, body weight,and carcass parameters of Sidama goats

A digestibility, feed intake, and carcass evaluation experiment using 20 yearling intact male Sidama goats weighing 16.4 ± 0.63 kg (mean ± SD) was conducted in Ethiopia with the objectives to determine feed intake, digestibility, body weight (BW) gain, and carcass parameters. The treatments included feeding natural pasture hay (T1, control) and supplementation with cottonseed cake (284 g—T2), linseed cake (250 g—T3), and noug seed cake (296 g—T4) on dry matter (DM) basis to supply 85 g crude protein (CP) per head per day. Randomized complete block design for feed intake and BW parameters and complete randomized design for digestibility and carcass parameters were used. Hay DM intake was higher (P < 0.01) for T1 than for the other treatments. T3 promoted higher (P < 0.01) DM (29.3 g/kg W^0.75/day) and CP (14.1 g/kg W^0.75/day) intake than T4 (8.9 g/kg W^0.75/day DM and 4.1 g/kg W^0.75/day CP). T3 showed better (P < 0.05) organic matter and CP digestibility than T2. Goats in T3 had higher nitrogen intake (P < 0.01) and retention (P < 0.05) than those in T1. Goats in T2 and T3 showed higher (P < 0.05) daily BW gain and final BW than those in T4 and T1. Goats in T2 and T3 had higher (P < 0.05) slaughter weight, empty BW, hot carcass weight, rib-eye muscle area, and dressing percentage on slaughter weight basis than those in T1. The results showed that T2 and T3 had similar effect on CP intake, daily BW gain, and carcass parameters for growing Sidama goats fed natural pasture hay.     

Effect of lipopolysaccharide on intranasal administration of liposomal Newcastle disease virus vaccine to SPF chickens

In order to potentiate the low immunogenicity of the inactivated Newcastle disease virus immunized into chickens by mucosal route, liposomes as a drug delivery system and LPS (lipopolysaccharide) as an immuno-stimulator were evaluated. Here, we report a new nasal delivery system of inactivated Newcastle disease virus (NDV) vaccine. The intranasal vaccine was based on different lipids to form MLV (multi-lamellar vehicles) liposomes. The liposomes had combined carrier and adjuvant activities, which induced strong systemic (serum) and local (lung and nasal) humoral responses in SPF (specific-pathogen-free) chickens, and provided protective immunity. PC-Lip (phosphatidylcholine-liposome) elicited significant mucosal secretary immunoglobulin A (s-IgA) levels (p < 0.05) in tracheal lavage fluid and serum IgG levels (p < 0.05). In response to virulent viral challenge, birds treated with PBS (phosphate buffered saline) as control group died, whereas 80% of chickens which received PC-Lip, PC-Lip-LPS, PS-Lip (phosphatidylserine-liposome), and PS-Lip-LPS survived. HAI titers were 1:2560 in the PS-Lip-LPS group and 1:1280 in the PC-Lip, PC-Lip-LPS, and PS-Lip groups after two vaccinations. The results suggest that PC-Lip or PS-Lip might thus be suitable as a potential adjuvant for mucosal vaccination against NDV in chickens.     

Immune responses to haemorrhagic septicaemia (HS) vaccination in <Emphasis Type="Italic">Trypanosoma evansi</Emphasis> infected buffalo-calves

To assess the immunosuppressive effect of Trypanosoma evansi infection in buffalo-calves on immune responses to heterologous antigen, the study was planned to examine the responses of haemorrhagic septicaemia vaccination in simultaneously and previously (80 days before vaccination) T. evansi-infected buffalo-calves. Eight buffalo-calves were divided into three groups. Buffalo-calves of group A (n = 3) were previously (80 days before primary vaccination with haemorrhagic septicaemia [HS] vaccine) infected with T. evansi (1 × 10⁷ tryps.calf⁻¹; sc) and that of group B (n = 3) were infected with T. evansi (1 × 10⁷ tryps.calf⁻¹; sc) on the day of primary vaccination with HS vaccine. Two healthy uninfected control calves given only HS vaccine were kept in group C. All the buffalo-calves were given a booster dose of vaccine 21 days post-primary vaccination (PPV). Twenty eight days PPV, animals of group A were given trypanocidal quinapyramine prosalt at 6.66 mg kg⁻¹. Immunosuppressive effect of T. evansi infection was evident from day 7 PPV with HS vaccine. The effect was more pronounced in previously T. evansi-infected buffalo-calves as compared with simultaneously infected buffalo-calves. Group A buffalo-calves appeared to have recovered from the immunosuppressive effect after 28 days post-trypanocidal treatment as observed by humoral and cell-mediated immune responses. Immunosuppressive effect to HS vaccination was observed in T. evansi-infected buffalo-calves, and trypanocidal therapy enabled the calves to mount the responses similar to uninfected controls.     

<Emphasis Type="Italic">Oestrus ovis</Emphasis> larval myiasis among sheep and goats in Central Oromia,Ethiopia

A study was conducted to determine the prevalence, larval burden, and associated gross pathological lesions of Oestrus ovis in sheep and goats slaughtered at Luna export abattoir in Central Oromia from November 2007 to March 2008. For this purpose, a total of heads of 431 goats and 369 sheep were thoroughly examined for the presence of first (L1), second (L2), and third (L3) larval stages according to standard procedures. O. ovis larvae were detected in 349(94.6%) sheep and 381(88.4%) goats. All three larval instars were observed in each study months. Statistically significant variation (χ ² = 29.2676, df = 6, P < 0.05) was observed in the prevalence of O. ovis among small ruminants of different origins. Likewise, statistically significant (χ ² = 68.3, df = 4, P < 0.05) difference was recorded in the prevalence of O. ovis in sheep and goats among different study months. The overall monthly prevalence ranged from 77.7% in November to 98.8% in March. The prevalence of O. ovis in small ruminants of less than 1 year of age was significantly (χ ² = 8, df = 1, P < 0.05) higher than those with greater than 1 year of age. An overall proportion of 33.8%, 40.1%, and 26.1% were recorded for L1, L2 and L3, respectively. Whereas 6.8 monthly mean larval burden per individual infested animal was noticed. Out of the total infested heads in goats, 33.6% had catarrhal discharges, 16.8% purulent exudates, 64.83% rhinitis, 68.77% sinusitis, 14.2% pharyngitis, and 9.2% bloody exudates. Similarly, of the total infested heads of sheep, 18.9% purulent exudates, 80.8% rhinitis, 71.9% sinusitis, 13.5% pharyngitis, and 7.7% bloody exudates gross lesions were recorded.     

Pathophysiological and immune cell responses in calves prior to and following lung challenge with formalin-killed Pasteurella multocida biotype A:3 and protection studies involving subsequent homologous live challenge

Pneumonic pasteurellosis is a common respiratory infection in cattle that has major economic and welfare implications world-wide and the incidence in the UK due to Pasteurella multocida, currently the same as that associated with Mannheimia haemolytica, is increasing. Whereas much is known regarding the pathogenesis of M. haemolytica infections little information is available on the pathogenic process of pasteurellosis initiated by P. multocida. In the present work calf systemic and innate immune responses to intratracheal challenge with formalin-killed P. multocida biotype A:3 and to subsequent experimental lung infection with live P. multocida were investigated. Eight-week-old calves were challenged intratracheally on day 0 with either 10⁹ colony forming units (cfu) of formalin-killed P. multocida biotype A:3 in 300 ml saline (n=10) or 300 ml saline alone (n=10), followed, at day 21, by challenge with 10⁹ cfu live P. multocida. Pathophysiological and lung phagocyte responses were assessed by clinical monitoring, sequential lung lavage and blood sampling. Results for samples obtained before, during and after challenge showed clinical and acute phase protein responses to both bacterial culture and saline control treatments, although higher responses were associated with bacterial challenge. Phagocytosis of P. multocida during 1 h incubation periods with lavaged cells in vitro was unaffected by exposure in vivo to killed P. multocida and there was evidence that P. multocida was able to survive intracellularly during this assay. There was no indication that lung exposure to formalin-killed P. multocida conferred protection against subsequent homologous live challenge.     

Huizache (<Emphasis Type="Italic">Acacia farnesiana</Emphasis>) whole pods (flesh and seeds) as an alternative feed for sheep in Mexico

Two experiments were undertaken to evaluate the use of pods from Huizache (Acacia farnesiana), common in the arid and semiarid regions of Mexico, on the perfromance and apparent digestibility in Pelibuey Mexican hair growing ewe lambs. Twenty-four Pelibuey ewe lambs were used in the animal performance experiment, with a mean live weight of 14.91 ± 1.48 kg, randomnly allocated to three groups which received ad libitum for 77 days (11 weeks) experimental whole rations T0 with 0%, T12 with 12% or T24 with 24% inclusión of dried and ground Huizache pods. Dry matter intakes (g/kg ^0.75 daily) were 83, 95, 90 for T0, T12, and T24 respectively (P > 0.05). Mean daily live-weight gain was 90, 75, and 63 g/day for T0, T12, and T24 (P < 0.001). Nine Pelibuey ewe lambs were used to determine apparent digestibility in vivo of the experimental diets using a 3 × 3 latin square design repeated three times. There were differences in the digestibility of dry matter (P < 0.001), organic matter (P < 0.001), nitrogen (P < 0.031), neutral detergent fibre (P < 0.002), and acid detergent fibre (P < 0.001) being lower in T24. Huizache pods may be an alternative feed when included up to 12% of dry matter in the diets for sheep growing moderately.     

Nasal immunization with major epitope-containing ApxIIA toxin fragment induces protective immunity against challenge infection with Actinobacillus pleuropneumoniae in a murine model

Actinobacillus pleuropneumoniae is an infective agent that leads to porcine pleuropneumonia, a disease that causes severe economic losses in the swine industry. Based on the fact that the respiratory tract is the primary site for bacterial infection, it has been suggested that bacterial exclusion in the respiratory tract through mucosal immune induction is the most effective disease prevention strategy. ApxIIA is a vaccine candidate against A. pleuropneumoniae infection, and fragment #5 (aa. 439–801) of ApxIIA contains the major epitopes for effective vaccination. In this study, we used mice to verify the efficacy of intranasal immunization with fragment #5 in the induction of protective immunity against nasal challenge with A. pleuropneumoniae and compared its efficacy with that of subcutaneous immunization. Intranasal immunization of the fragment induced significantly higher systemic and mucosal immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. Intranasal immunization not only efficiently inhibited the bacterial colonization in respiratory organs, but also prevented alveolar tissue damage in infectious condition similar to that of a contaminated pig. Moreover, intranasal immunization with fragment #5 provided acquired protective immunity against intranasal challenge with A. pleuropneumoniae serotype 2. In addition, it conferred cross-protection against serotype 5, a heterologous pathogen that causes severe disease by ApxI and ApxII secretion. Collectively, intranasal immunization with fragment #5 of ApxIIA can be considered an efficient protective immunization procedure against A. pleuropneumoniae infection.