Paratuberculosis Research Articles

Johne's Disease Update

Mycobacterium paratuberculosis causes a granulomatous enteritis in cattle that is commonly known as Johne's Disease or paratuberculosis. Young calves acquire the infection in utero or within the first few months of age by the fecal-oral route. Available diagnostic tests have limitations, but when used with appropriate management practices, a control/eradication program can be designed to meet the needs of the producer. Control programs are aimed at minimizing or eliminating exposure to the organism. There is no curative treatment for paratuberculosis.

A preliminary study investigating effects of oral monensin sodium in an enteric Mycobacterium avium ssp. paratuberculosis infection model of calves

Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of Johne's disease, an enteric infection of ruminants that causes significant economic burden for dairy and beef producers. Efforts to control MAP in endemic herds typically focus on herd management practices such as limiting exposure or early culling of infected animals and, occasionally, vaccination. The ionophore monensin sodium may have protective effects against MAP both in vivo and in vitro; however, this has not been thoroughly evaluated experimentally. Using a direct intestinal MAP challenge model, we have observed similarities regarding persistence of MAP in tissues and apparent resilience to infection compared with experimental oral infection or natural disease. Here we sought to investigate the effects of oral monensin supplementation in experimentally MAP-infected calves. We examined the persistence of MAP in the intestinal tissues, MAP-induced intestinal inflammation, fecal MAP shedding, and seroconversion using a commercial serologic assay. Monensin-supplemented MAP-infected calves demonstrated evidence for resilience to MAP infection earlier in this study compared with monensin-free MAP-infected calves. However, statistical modeling did not identify a significant effect of monensin on outcomes of infection, and more work is required to understand how monensin affects early tissue colonization of MAP in calves.

Evaluation of newly developed ‘six recombinant secretary proteins based ‘cocktail ELISA’ and ‘whole cell lysate’ based ‘indigenous ELISA’ and tissue microscopy’ with ‘Gold standard’ histo-pathology for the diagnosis of Johne’s disease in slaughtered goats and buffaloes

Three screening tests {(newly developed, six recombinant secretory proteins based 'cocktail ELISA', in-house robust 'indigenous ELISA' based on semi-purified protoplasmic antigens and tissue microscopy were evaluated with 'Gold standard', histo-pathology for the diagnosis of Johne's disease in goats and buffaloes. Serum and tissues {mesenteric lymph nodes and intestines) were driven from farmer's goats (n = 77) and buffaloes (n = 40) slaughtered for harvesting meat and farm goats (n = 77), died and necropsied. Twenty seven (35%) goats and 23 (57.5%) buffaloes were positive in all the four tests. Of 134 tissues screened by histo-pathology, 79.8% MLN and 76.8%, intestines, were positive for MAP infection. In tissue microscopy, 55.2 and 52.3%, goats and buffaloes were positive, respectively. Of 117 sera screened by i_ELISA, 58.4 and 70.0%, goats and buffaloes were positive, respectively. Whereas, c_ELISA detected 55.8 and 62.5%, goats and buffaloes, positives, respectively. Twelve tissues (70.5%) of goats necropsied were positive, both in tissue microscopy and histo-pathology. Most significant gross findings were serous atrophy of the fat and mild to moderate, diffuse thickening of terminal ileum, especially at ileo-caecal junction with or without transverse / longitudinal corrugations. In histo-pathology grade III and IV lesions were significantly low as compared to grade I and II. Of the four tests used for screening 268 samples, histo-pathology was most sensitive (78.3%), followed by i_ELISA (62.3%), c_ELISA (58.9%) and tissue microscopy (58.9%). Between two ELISA tests, c_ELISA using six recombinants secretory proteins, had higher specificity as compared to i_ELISA.

An update on Mycobacterium avium subspecies paratuberculosis antigens and their role in the diagnosis of Johne's disease.

Mycobacterium avium subsp. paratuberculosis (MAP) is responsible for Johne's disease (JD) or paratuberculosis. Diagnosis of MAP infection by measuring host cell-mediated and humoral immune responses has been a major focus in MAP research. For this purpose, several MAP antigens such as secreted protein, cell envelope protein, cell-mediated immune and lipoprotein antigens have been identified and tested to measure their diagnostic utility with varying degree of success. Identifying the optimal antigen or antigen combinations for diagnosis of infected animals is hindered by the complex nature of the disease, prolonged subclinical infection, the differential expression of antigens and scarcity of well characterized MAP-specific epitopes making selection of a single MAP antigen very difficult. Thus, multiplexing of antigens with larger scale and longitudinal studies may lead to development of cost-effective next generation serodiagnostics. This mini review focuses on the role of different MAP antigens in the diagnosis of JD.

The specificity of phage testing for MAP — where might it fit into the diagnostic armoury?

The current individual tools available for the diagnosis of Johne's disease are far from suitable to tackle this endemic disease. Culture, polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) tests, when used together can be useful in managing the disease in the later stages of infection at a herd level. They are, however, ill-suited to detecting the causative agent Mycobacterium avium subsp. paratuberculosis (MAP) at the early stages of infection and at an individual level. Phage technology offers another tool in the attempt to better manage and control this disease. Phage-technology has been demonstrated to rapidly and sensitively detect and specifically identify viable MAP in the milk and blood of cattle. Although in relatively-early stages of development phage technology offers a strong addition to the armoury of tests used to detect MAP in blood and milk, and may go on to be part of ongoing control measures to reduce the burden of disease to farmers and veterinarians.

Epidemiology and risk factors of Mycobacterium avium subspecies paratuberculosis in semi-extensive dairy sheep and goat farms of Apulia, southern Italy

Paratuberculosis (PTB) is a well-documented chronic and sometimes fatal infection that affects the small intestine of ruminants both in captive and free-ranging living conditions. Although the infection has been detected on small ruminant farms worldwide, epidemiological and risk factor information regarding Mycobacterium avium subspecies paratuberculosis (MAP) infection on semi-extensive sheep and goat farms is generally still scarce, particularly in the Mediterranean area and in Italy. This paper reports the epidemiological findings and risk factors of the infection on semi-extensive sheep and goat farms in Apulia. It particularly focuses on the involvement of biological, structural and management factors, as well as the farmers’ socioeconomic data as predisposing causes of PTB, and investigates their possible connection with the survivability of the infection.The true seroprevalence values of MAP on semi-extensive Apulian sheep and goat farms are reported and the risk factors causing the spread of infection analyzed. Data were collected through a two-year survey over the whole regional area, involving 419 farms, 16,903 sheep and 9369 goats. The epidemiological results showed a true seroprevalence of 66.2% for flocks and of 9.7% at the animal level. Analysis of the risk factors showed that the spread of infection occurs via several concomitant biological, managerial, and farmer-related factors.The survey highlighted the need for urgent and suitable control plans providing guidelines to help Apulian farmers and their veterinarians contain and eliminate the disease, prevent biological risks for animal and humans as well as negative economic effects on the regional livestock sector.

Mycobacterium tuberculosis extracellular vesicle-associated lipoprotein LpqH as a potential biomarker to distinguish paratuberculosis infection or vaccination from tuberculosis infection

BackgroundBoth bovine tuberculosis (TB) and paratuberculosis (PTB) are serious and widespread bacterial infections affecting many domestic and wild animal species. However, current vaccines do not confer complete protection and cause interference with other diagnostics tests, including bovine TB. Therefore, the development of “Differentiating Infected from Vaccinated Animals” (DIVA) tests are a pressing need. In this study, we have tested the feasibility of mycobacterial extracellular vesicles (EVs) as potential source of biomarkers to discriminate between Mycobacterium bovis infected, Mycobacterium avium subsp. paratuberculosis (MAP) infected and MAP-vaccinated cows. We have, initially, characterized vesicle production in the two most medically relevant species of mycobacteria for livestock, MAP and M. bovis, for being responsible for tuberculosis (TB) and paratuberculosis (PTB).ResultsOur results indicate that these two species produce EVs with different kinetics, morphology and size distribution. Analysis of the immunogenicity of both type of EVs showed some cross reactivity with sera from PTB+ and TB+ cows, suggesting a limited diagnostic capacity for both EVs. Conversely, we noticed that Mycobacterium tuberculosis (Mtb) EVs showed some differential reactivity between sera from MAP-vaccinated or PTB+ cows from TB+ ones. Mass spectrometry analysis (MS) identified a 19-kDa EV-associated lipoprotein as the main source of the differential reactivity.ConclusionsLpqH could be a good plasma biomarker with capacity to distinguish PTB+ or MAP-vaccinated cows from cows infected with TB.

Development and validation of a loop-mediated isothermal amplification assay-a rapid and sensitive detection tool for Mycobacterium avium subsp. paratuberculosis in small ruminants.

The aim of this study was to design an assay for the identification of Mycobacterium avium subsp. paratuberculosis (MAP) to be used in faeces and milk samples of small ruminants with a loop-mediated isothermal amplification (LAMP) system, as a time-saving and user-friendly method in contrast to real-time PCR. For the detection of MAP in milk and faeces of small ruminants, we developed a set of primers, specific for the target gene ISMap02. The analytical sensitivity of LAMP, when targeting ISMap02, showed a DNA detection limit of 10fgμl-1 . After performing spiking experiments with two MAP reference strains, DSM 44133 and ATCC 19698T , the limit of detection, using the LAMP protocol described herein were 3·8 MAP CFU per ml milk and 12·5 MAP CFU per gram faeces. All LAMP results during the establishment of the assay were compared to those of the real-time PCR results. An internal amplification control was incorporated into the assay to exclude false-negative results produced and had no significant negative impact on the analytical sensitivity. Validation of the assay was confirmed by testing field samples of faeces and revising the results with real-time PCR. Our study conducted the first MAP detection system with a LAMP targeting ISMap02. Due to the positive results we encourage the use of LAMP in combination with ISMap02, when detecting MAP in faeces samples, as an alternative to targeting other genes as f57 or IS900. Further research on MAP detection in different matrices like raw milk, tissue or sperm with this system is recommended. This study provides new achievements in MAP diagnostic. Especially small ruminants do not show signs of diarrhoea until the terminal stage of the illness. The greatest task in fighting MAP is to rule out animals, which shed MAP with faeces and milk before showing symptoms of Johne's disease. Worldwide there is a need to eradicate animals, which are low MAP shedders to stop the illness spreading in animal holdings. MAP detection with LAMP is time saving, easy to use, does not need expensive equipment, as, for example, PCR kits and can be used without access to laboratories. The target gene ISMap02 was shown to be a specific insertion element for MAP and is a reliable aim in future MAP detection studies.

Molecular and bacteriological investigation of Johne’s disease in dairy cattle

Johne's disease (JD) caused by Mycobacterium avium subsp. paratuberculosis (MAP) represents a real threat to the agriculture and dairy food industries and believed to be a potential public health problem. No treatment for the disease exists and controlling the disease is difficult due to its long latent period. This study aimed to evaluate two diagnostic methods culture and PCR assay for detection of MAP in feces under the Egyptian circumstances. A total 200 fecal samples were collected from dairy cattle showed chronic diarrhea and have history of Johne’s infection. Fecal culturing revealed 87 (43.5 %) positive from the total collected samples. In another side, the PCR targeting IS1311 revealed that 94 (47%) out of 200 of examined cattle were positive for MAP. Consequently, the both tests showed good agreement between them (Kappa value = 0.869). The sequence analysis of the obtained isolate showed that the Egyptian isolates (LC428286 and LC428285) have high similarity with MAP strain JII-1961 from Germany and MAP 316V strain of Netherlands reach up to 98%.

The immunogenicity and tissue reactivity of Mycobacterium avium subsp paratuberculosis inactivated whole cell vaccine is dependent on the adjuvant used

Johne's disease (JD) is a chronic enteritis caused by Mycobacterium avium subspecies paratuberculosis (MAP). Current commercial vaccines are effective in reducing the occurrence of clinical disease although vaccinated animals can still become infected and transmit MAP. Many vaccinated sheep develop severe injection site lesions. In this study a range of adjuvants (MontanideTM ISA 50V, ISA 50V2, ISA 61VG, ISA 70 M VG, ISA 71 VG, ISA 201 VG and Gel 01 PR) formulated with heat-killed MAP were tested to determine the incidence of injection site lesions and the types of immune profiles generated in sheep. All the novel formulations produced fewer injection site lesions than a commercial vaccine (Gudair®). The immune profiles of the sheep differed between treatment groups, with the strength of the antibody and cell mediated immune responses being dependant on the adjuvant used. One of the novel vaccines resulted in a reduced IFN-γ immune response when a second “booster” dose was administered. These findings have significance for JD vaccine development because it may be possible to uncouple protective immunity from excessive tissue reactivity, and apparently poorly immunogenic antigens may be re-examined to determine if an appropriate immune profile can be established using different adjuvants. It may also be possible to formulate vaccines that produce targeted immunological profiles suited to protection against other pathogens, i.e. those for which a bias towards cellular or humoral immunity would be advantageous based on understanding of pathogenesis.

Global scenario of paratuberculosis: a threat to livestock sector

ABSTRACT Mycobacterium avium subspecies paratuberculosis (MAP) implicated in paratuberculosis (PTB) infection in animals and Crohn’s disease in human poses enormous economic loss to the farmers. The noticeable withstand ability of organism to environment, faecal shedding of organism during subclinical and clinical infection consequence frequent outbreak. The obvious view of geographical distribution of PTB all over the world is needed in order to manage the disease outbreaks in a prompt manner, implementation of disease control programme in an efficient way and also for international smooth trading of animals without any further outbreak. Global scenario of PTB has been concluded in the present context on the basis of available previous prevalence studies of PTB in different continents. Prevalence of PTB in Africa (53%), India (23.3%), Europe (20%), South America (18.3%), North America (16.9%) and Australia (6.8%) has been documented. This review tries to bring out a clear picture about the prevalence of PTB throughout the world in domesticated animals which may help in understanding the geographical distribution of the disease according to its epidemiological importance and minimizing the endemicity of disease.

Invited review: A systematic review of the effects of early separation on dairy cow and calf health

Concern from the public is growing regarding early cow-calf separation, yet proponents of this practice maintain that artificial rearing is critical for cow and calf health. Early separation is assumed to reduce the risk of transfer of pathogens from dam to neonatal calf, but a wide range of health benefits associated with extended cow-calf contact has also been documented. The aim of this systematic review was to report and synthesize conclusions from the literature on dairy cow and calf health in conventional rearing versus cow-calf contact systems. Peer-reviewed, published manuscripts, written in English, directly comparing dairy cow or calf health in artificial versus suckling systems, were eligible for inclusion. We conducted 7 targeted searches using Web of Science to identify key literature on important health conditions. The resulting manuscripts underwent a 4-step appraisal process, and further manuscripts were sourced from reference lists. This process resulted in a final sample of 70 articles that addressed cow and calf health. Sufficient literature was available to assess mastitis in cows, and scours, cryptosporidiosis, Johne's disease, pneumonia, immunity, and mortality in calves. The results for cryptosporidiosis, pneumonia, immunity, and mortality were mixed, with some differences between studies likely attributable to flawed comparisons between cohorts. Overall, the articles addressing calf scours and mastitis pointed to beneficial or no effects of suckling. The studies addressing Johne's disease did not find cow-calf contact to be a significant risk factor. In conclusion, the scientific peer-reviewed literature on cow and calf health provides no consistent evidence in support of early separation.

Modeling the effects of infection status and hygiene practices on Mycobacterium avium subspecies paratuberculosis contamination in bulk tank milk

Infectious diseases in dairy cattle are of significant concern to dairy industries because of their huge impact on animal health, milk production, and economics. Mycobacterium avium subsp. paratuberculosis (MAP) is a pathogenic bacterium that causes Johne's disease, one of the important endemic infectious diseases in dairy cattle. Contamination of bulk tank milk with MAP can occur through direct shedding into milk by infected cows (internal route), fecal contamination (fecal route), or introduction of soil and water containing MAP (environmental route). Humans can be exposed to MAP via raw milk consumption; additionally, there are reports of MAP survival in milk after pasteurization. The risk of human consumption is particularly important due to an association between MAP and human Crohn's disease. In the current study, we used a probabilistic modeling framework to predict the level of MAP contamination per liter in the bulk tank milk and weigh the relative importance of each contamination route. Our model focused on several different infection statuses and the contribution of each group to environmental and fecal contamination, in addition to internal route shedding. We assessed the influence of common hygiene practices, such as washing of udders before milking and the use of milk filters, on the concentration of MAP in bulk tank milk. We extracted parameters and their distributions from national surveys and thorough literature search. Our baseline model comprising all hygiene practices provided an average estimate of 0.76 log CFU/L for the final concentration of MAP in bulk tank milk, with a maximum of 6.70 log CFU/L and a minimum of 0.04 log CFU/L depending on herd size and the ratio of infection statuses. Results from sensitivity analyses indicated that the average fecal contamination showed the greatest impact on the final MAP concentration per liter in bulk tank milk, followed by herd size and washing efficiency. This study emphasized that good hygiene practices are crucial for maintaining the quality of raw milk in an endemically-infected dairy herd.

A peptide-based vaccine for Mycobacterium avium subspecies paratuberculosis

Recent efforts to develop a live attenuated vaccine against Mycobacterium avium subsp. paratuberculosis (Map), the causative agent of Johne’s disease (JD), revealed relA is important in Map virulence. Deletion of the relA gene impairs the ability of Map to establish a persistent infection. Analysis of the basis for this observation revealed infection with a relA deletion mutant (ΔrelA) elicits development of cytotoxic CD8 T cells (CTL) with the ability to kill intracellular bacteria. Further analysis of the recall response elicited by ΔrelA vaccination showed a 35 kDa membrane peptide (MMP) is one of the targets of the immune response, suggesting it might be possible to develop a peptide-based vaccine based on MMP. To explore this possibility, ex vivo vaccination studies were conducted with MMP alone and incorporated into a nanoparticle (NP) vector comprised of poly (D, L-lactide-co-glycolide) and monophosphoryl lipid A (PLGA/MPLA). As reported, ex vivo vaccination studies showed CD8 CTL were elicited with classic and monocyte derived dendritic cells (cDC and MoDC) pulsed with MMP alone and incorporated into a PGLA/MPLA vector. Incorporation of MMP into a NP vector enhanced the ability of CD8 CTL to kill intracellular bacteria. The findings indicate incorporation of MMP into a PGLA/MPLA nanoparticle vector is one of the possible ways to develop a MMP based vaccine for Johne’s disease.

The inhibitory effect of nisin on Mycobacterium avium ssp. paratuberculosis and its effect on mycobacterial cell wall

Infection with Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) is a widespread problem in the United States and worldwide, and it constitutes a significant health problem for dairy animals with a potential effect on human health. Mycobacterium paratuberculosis is easily transmitted through consumption of contaminated milk; therefore, finding safe methods to reduce the mycobacterial load in milk and other dairy products is important to the dairy industry. The main objective of the current study was to investigate the effect of natural products, such as bacteriocins designated as "generally regarded as safe" (GRAS), on the survival of M. paratuberculosis in milk. Commercially synthesized bacteriocin (nisin) was used to examine its effect on the survival of laboratory and field isolates of M. paratuberculosis and in contaminated milk. Surprisingly, nisin had a higher minimum inhibitory concentration (MIC) against the laboratory strain (M. paratuberculosis K10), at 500 U/mL, than against field isolates (e.g., M. paratuberculosis 4B and JTC 1281), at 15 U/mL. In milk, growth of M. paratuberculosis was inhibited after treatment with levels of nisin that are permissible in human food at 4°C and 37°C. Using both fluorescent and scanning electron microscopy, we were able to identify defects in the bacterial cell walls of treated cultures. Our analysis indicated that nisin reduced membrane integrity by forming pores in the mycobacterial cell wall, thereby decreasing survival of M. paratuberculosis. Thus, nisin treatment of milk could be implemented as a control measure to reduce M. paratuberculosis secreted in milk from infected herds. Nisin could also be used to reduce M. paratuberculosis in colostrum given to calves from infected animals, improving biosecurity control in dairy herds affected by Johne's disease.

Genetic association of polymorphisms in bovine TLR2 and TLR4 genes with Mycobacterium avium subspecies paratuberculosis infection in Indian cattle population.

Toll like receptors (TLRs) are pattern recognition molecules involved in cellular recognition of Mycobacterium avium subspecies paratuberculosis (MAP), the infectious agent causing Paratuberculosis (PTB), a notified disease of domestic and wild ruminants. The present study was undertaken to investigate the presence of single nucleotide polymorphisms (SNPs) in TLR2 and TLR4 gene and to evaluate association of these SNPs with occurrence of PTB in Indian cattle. A total of 213 cattle, were subjected to multiple diagnostic tests viz. Johnin PPD, ELISA test (Indigenous and Parachek kit method), fecal microscopy and fecal culture for detection of MAP infection. Based on screening results 51 animals each were assigned to case and control population. Two SNPs viz. rs55617172, rs41830058 in TLR2 gene and two SNPs viz. rs8193046, rs8193060 in TLR4 gene and were genotyped by PCR-RFLP method. All SNPs were found to be polymorphic except rs41830058 in the case-control population. Both SNPs in TLR4 gene but none in TLR2 genes were significantly associated with the occurrence of PTB in our population. The genotypes in SNP rs8193046 and SNP rs8193060 were significantly (P < 0.01) different in case-control population. These findings suggest that SNPs rs8193046 and rs8193060 are likely a potential marker against MAP infection and a selection programme eliminating AG genotype for rs8193046 and CT genotype for rs8193060 might be beneficial in conferring resistance to MAP infection in Indian cattle population.

Complete Genome Sequence of the Telford Type S Strain of Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease (JD). Here, we report the complete genome sequence of Telford 9.2, a well-characterized representative strain of the M. avium subsp. paratuberculosis S subtype that is endemic in New Zealand and Australian sheep.

Exploring dairy producer and veterinarian perceptions of barriers and motivators to adopting on-farm management practices for Johne's disease control in Ontario, Canada

Motivating uptake of management change recommendations requires knowledge of the barriers and motivators influencing producer behavior. This study investigated dairy producers' and veterinarians' perceptions of the barriers and motivators influencing the adoption of Johne's disease (JD) control recommendations in Ontario, Canada. Eight focus groups, 6 with dairy producers and 2 with veterinarians, were conducted and thematically analyzed. Both producer and veterinarian groups identified physical resources (i.e., time, money, infrastructure) and producer mindset (i.e., perceived priority of JD, perceived practicality of JD control recommendations) as key barriers to adoption. Producers tended to prioritize JD control on their farm based on their lived experiences with JD and their view of the public's concern about JD. Many agreed that JD recommendations should focus on biosecurity more holistically and emphasize the broader health benefits of limiting calf exposure to many fecal-orally transmitted diseases. Producers also highlighted that some recommendations for on-farm change (i.e., keeping a closed herd, buying from low-risk herds) were unrealistic or too difficult to perform and often disrupted their habits or routine. In contrast, veterinarians suggested that most recommendations were practical and are routinely recommended. Participants suggested both extrinsic (i.e., incentives, premiums, penalties and regulations, and extension and communication) and intrinsic (i.e., pride and responsibility) methods for motivating producers. This study highlights the importance of producer mindset in on-farm change and offers insights into the attitudes and perceived barriers influencing on-farm change.

Candidate genes associated with the heritable humoral response to Mycobacterium avium ssp. paratuberculosis in dairy cows have factors in common with gastrointestinal diseases in humans

Infection of cattle with bovine paratuberculosis (i.e., Johne's disease) is caused by Mycobacterium avium ssp. paratuberculosis (MAP) and results in a chronic incurable gastroenteritis. This disease, which has economic ramifications for the cattle industry, is increasing in detected prevalence globally; subclinically infected animals can silently shed the bacterium into the environment for years, exposing contemporaries and hampering disease-control programs. The objective of the present study was to first quantify the genetic parameters for humoral response to MAP in dairy cattle. This was followed by a genome-based association analysis and subsequent downstream bioinformatic analyses from imputed whole genome sequence SNP data. After edits, ELISA test records were available on 136,767 cows; analyses were also undertaken on a subset of 33,818 of these animals from herds with at least 5 MAP ELISA-positive cows, with at least 1 of those positive cows being homebred. Variance components were estimated using univariate animal and sire linear mixed models. The heritability calculated from the animal model for humoral response to MAP using alternative phenotype definitions varied from 0.02 (standard error = 0.003) to 0.05 (standard error = 0.008). The genome-based associations were undertaken within a mixed model framework using weighted deregressed estimated breeding values as a dependent variable on 1,883 phenotyped animals that were ≥87.5% Holstein-Friesian. Putative susceptibility quantitative trait loci (QTL) were identified on Bos taurus autosome 1, 3, 5, 6, 8, 9, 10, 11, 13, 14, 18, 21, 23, 25, 26, 27, and 29; mapping the most significant SNP to genes within and overlapping these QTL revealed that the most significant associations were with the 10 functional candidate genes KALRN, ZBTB20, LPP, SLA2, FI3A1, LRCH3, DNAJC6, ZDHHC14, SNX1, and HAS2. Pathway analysis failed to reveal significantly enriched biological pathways, when both bovine-specific pathway data and human ortholog data were taken into account. The existence of genetic variation for MAP susceptibility in a large data set of dairy cows signifies the potential of breeding programs for reducing MAP susceptibility. Furthermore, the identification of susceptible QTL facilitates greater biological understanding of bovine paratuberculosis and potential therapeutic targets for future investigation. The novel molecular similarities identified between bovine paratuberculosis and human inflammatory bowel disease suggest potential for human therapeutic interventions to be translated to veterinary medicine and vice versa.

Mycobacterium avium subspecies paratuberculosis is able to manipulate host lipid metabolism and accumulate cholesterol within macrophages

Johne's disease is a chronic wasting disease of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP). Closely related pathogenic mycobacteria such as M. tuberculosis are capable of altering host lipid metabolism, highlighting the need to explore the role of lipid metabolism contributing to intracellular survival. This study aimed to identify whether MAP is able to manipulate host lipid metabolic pathways and accumulate host cholesterol during early infection. Macrophages were exposed to four different MAP strains and non-pathogenic M. phlei for up to 72 h, with changes to lipid metabolism examined using fluorescent microscopy and gene expression. MAP-infected macrophages displayed strain-dependent differences to intracellular cholesterol levels during early infection, however showed similarly increased intracellular cholesterol at later timepoints. Gene expression revealed that MAP strains similarly activate the host immune response in a conserved manner compared to M. phlei. MAP significantly upregulated host genes associated with lipid efflux and endocytosis. Moreover, lipid biosynthesis genes were differentially regulated in a strain-dependent manner following MAP infection. Collectively, these results demonstrate that MAP manipulates host lipid metabolism during early infection, however the extent of these modulations are strain-dependent. These findings reflect a conserved pathway contributing to intracellular MAP survival.