Mycoplasma (M.) hyopneumoniae is the main pathogen of porcine enzootic pneumonia (PEP). Its controlling is challenging, and requires alternative strategies. This study aimed to develop an oral vaccine against M. hyopneumoniae using a nanostructured mesoporous silica (SBA-15) as an adjuvant, and compare its effect with an intramuscular (IM) commercial vaccine (CV). Fifty 24 day-old M. hyopneumoniae-free piglets composed five equal groups for different immunization protocols, consisting of a CV and/or oral immunization (OI). Control piglets did not receive any form of immunization. All piglets were challenged with M. hyopneumoniae strain 232 on D49 by tracheal route. IgA antibody response in the respiratory tract, bacterial shedding and serum IgG were evaluated. The piglets were euthanized on 28 (D77) and 56 (D105) days post-infection. Lung lesions were macroscopically evaluated; lung fragments and bronchoalveolar fluid (BALF) were collected for estimation of bacterial loads by qPCR and/or histopathology examination. All immunization protocols induced reduction on Mycoplasma-like macroscopic lung lesions. IgA Ab responses anti-M. hyopneumoniae, the expression of IL-4 cytokine and a lower expression of IL-8 were induced by CV and OI vaccines, while IgG was induced only by CV. Oral immunization using silica as a carrier-adjuvant can be viable in controlling M. hyopneumoniae infection.
Background So far, three porcine hemoplasmas (PH) have been identified, namely Mycoplasma suis, Mycoplasma parvum, and Mycoplasma haemosuis. The first one is the main agent associated with porcine hemoplasmosis, a possible cause of economic losses in pig production. Thus, this work aimed to detect and quantify PH 16S rRNA in finishing pigs and to associate its load estimate with average daily weight gain (ADWG). For this purpose, whole blood samples from 318 pigs were collected at an age of 75 days (d0) when the pigs entered the finishing phase and 105 days later (d105). To calculate ADWG, the animals were weighed at the abovementioned dates. Then, DNA from blood samples were submitted to a qPCR targeting the 16S rRNA gene for PH. Spearman correlation test was performed to investigate potential associations between ADWG and the quantification values. Lastly, the molecular characterization of PH was done by sequencing the 23S rDNA gene. Results Out of the 318 samples, 190 (59.74%) were positive on d0, and 304 (95.6%) were positive on d105. A significant correlation was observed (p < 0.05), albeit with a low coefficient value (0.18), when comparing ADWG with quantification values on d105. The phylogenetic analysis based on the 23S rDNA gene showed that four sequences were closely related to M. parvum, and one sequence was positioned in the M. suis cluster. Conclusion Two PH, M. suis and M. parvum, were detected in a Brazilian pig farm. Moreover, increasing occurrence through time was observed, which may have affected the productive performance of positive animals, mainly at the end of the finishing phase, when antimicrobials are removed.
Porcine Respiratory Diseases Complex (PRDC) is a multifactorial disease that involves several bacterial pathogens, including Mycoplasma hyopneumoniae (M. hyopneumoniae), Actinobacillus pleuropneumoniae (A. pleuropneumoniae), Pasteurella multocida (P. multocida), Glaesserella parasuis (G. parasuis), and Streptococcus suis (S. suis). In pigs, the infection may cause lesions such pleurisy, which can lead to carcass condemnation. Hence, 1015 carcasses were selected from three different commercial pig farms, where the respiratory conditions were evaluated using slaughterhouse pleurisy evaluation system (SPES) and classified into five groups. In total, 106 pleural and lung fragments were collected for qPCR testing to identify the five abovementioned pathogens. A moderate correlation between the severity of the lesions and the presence of P. multocida (R = 0.38) and A. pleuropneumoniae (R = 0.28) was observed. Concerning the lung samples, the severity of the lesions was moderately correlated with the presence of P. multocida (R = 0.43) and M. hyopneumoniae (R = 0.35). Moreover, there was a strong correlation between the presence of P. multocida and M.hyopneumoniae in the pleura (R = 0.82). Finally, this approach may be a useful tool to identify and quantify causative agents of PRDC using qPCR, providing a comprehensive evaluation of its relevance, strength, and potential application in the field as a surveillance tool for veterinarians.
Mycoplasma hyopneumoniae is the primary agent of Swine Enzootic Pneumonia (SEP). Vaccines reduce the clinical manifestation of the disease but do not prevent infection. The present study aimed to evaluate the use of antimicrobial drugs to minimize the impact of M. hyopneumoniae. For this, 32 pregnant female pigs and their litters were selected and then followed from birth to slaughter. The study involved three experimental groups that received metaphylactic treatment with different protocols involving tilmicosin, valnemulin, tulathromycin, and a control group to compare the effect of treatments against M. hyopneumoniae infection throughout the phases. Performance data were recorded, and the piglets were evaluated for the occurrence of cough. Nasal swab and blood collection was conducted periodically to detect M. hyopneumoniae shedding and anti-M. hyopneumoniae IgG, respectively. At slaughter, the lungs of animals from all groups were evaluated, and samples were collected for histopathological examination and qPCR for M. hyopneumoniae detection. All protocols promoted a reduction in consolidation lung lesions when compared to the control group. Individuals treated with valnemulin showed increased performance results, lower mortality, and low bacterial load in the lung. The results are promising and may indicate an alternative in the strategic control of M. hyopneumoniae infection in pigs.
Mycoplasma hyopneumoniae is a difficult-to-control bacterium since commercial vaccines do not prevent colonization and excretion. The present study aimed to evaluate the performance of an orally administered vaccine composed of antigens extracted from Mycoplasma hyopneumoniae and incorporated into mesoporous silica (SBA-15), which has an adjuvant-carrier function, aiming to potentiate the action of the commercial intramuscular vaccine. A total of 60 piglets were divided into four groups (n = 15) submitted to different vaccination protocols as follows, Group 1: oral SBA15 + commercial vaccine at 24 days after weaning, G2: oral vaccine on the third day of life + vaccine commercial vaccine at 24 days, G3: commercial vaccine at 24 days, and G4: commercial vaccine + oral vaccine at 24 days. On the first day, the piglets were weighed and, from the third day onwards, submitted to blood collections for the detection and quantification of anti-Mycoplasma hyopneumoniae IgG. Nasal swabs were collected to monitor IgA by ELISA, and oropharyngeal swabs were used to assess the bacterial load by qPCR. Biological samples were collected periodically from the third day of life until the 73rd day. At 41 days of life, 15 individuals of the same age, experimentally challenged with an inoculum containing M. hyopneumoniae, were co-housed with the animals from groups (1 to 4) in a single pen to increase the infection pressure during the nursery period. At 73 days, all piglets were euthanized, and lungs were evaluated by collecting samples for estimation of bacterial load by qPCR. Quantitative data obtained from physical parameters and laboratory investigation were analyzed by performing parametric or non-parametric statistical tests. Results indicate that animals from G2 showed smaller affected lung areas compared to G3. Animals from G2 and G4 had a low prevalence of animals shedding M. hyopneumoniae at 61 days of age. Additionally, no correlation was observed between lung lesions and M. hyopneumoniae load in lung and BALF samples in animals that received the oral vaccine, while a strong correlation was observed in other groups. In the present study, evidence points to the effectiveness of the oral vaccine developed for controlling M. hyopneumoniae in pig production under field conditions.
Mycoplasma (M.) hyopneumoniae, the etiological agent of swine enzootic pneumonia, has been reported to increase the susceptibility to secondary infections and modulate the respiratory microbiota in infected pigs. However, no studies have assessed the influence of M. hyopneumoniae on the respiratory microbiota diversity under experimental conditions. Therefore, this study evaluated the impact of M. hyopneumoniae infection on the respiratory microbiota of experimentally infected swine over time. To accomplish this, 12 weaned pigs from a M. hyopneumoniae-free farm were divided into two groups: M. hyopneumoniae strain 232 infected (n = 8) and non-infected (n = 4). The first group received 10 mL of Friis medium containing 107 CCU/mL of M. hyopneumoniae while the control group received 10 mL of sterile Friis medium. Inoculation of both groups was performed intratracheally when the animals were 35 days old (d0). At 28 days post-inoculation (dpi) and 56 dpi, 4 infected animals plus 2 controls were humanely euthanized, and biopsy samples of nasal turbinates (NT) and bronchus-alveolar lavage fluid (BALF) samples were collected. The DNA was extracted from the individual samples, and each group had the samples pooled and submitted to next-generation sequencing. Taxonomic analysis, alpha and beta diversity indexes, weighted unifrac, and unweighted unifrac distances were calculated. A high relative frequency (99%) of M. hyopneumoniae in BALF samples from infected animals was observed with no significant variation between time points. The infection did not seem to alter the diversity and evenness of bacterial communities in NT, thus, M. hyopneumoniae relative frequency was low in NT pools from infected animals (28 dpi—0.83%; 56 dpi—0.89%). PCoA diagrams showed that BALF samples from infected pigs were grouped and far from the control samples, whereas NT from infected animals were not separated from the control. Under the present coditions, M. hyopneumoniae infection influenced the lower respiratory microbiota, which could contribute to the increased susceptibility of infected animals to respiratory infections.
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