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Introduction. Infectious keratoconjunctivitis is a significant ocular disease found in confined sheep. Little information about the aetiological agents and their antimicrobial susceptibility is available. Gap Statement. There is limited information on the aetiological agents involved in keratoconjunctivitis outbreaks in sheep. Aim. The present research aimed to determine the bacterial aetiological factors involved in an outbreak of infectious keratoconjunctivitis in confined lambs. Methodology. Ocular swabs were collected from 23 randomly selected lambs, which were classified into three groups according to the severity of the lesion: group I (N=6; no ocular involvement), group II (N=8; less severe injuries) and group III (N=9; more severe injuries). Isolation of aerobic bacteria and antimicrobial susceptibility tests were carried out. Molecular detection of Mollicutes was performed, and positive samples were tested to confirm the presence of the following species: Mycoplasma conjunctivae , Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri . Results. Moraxella sp. and Mollicutes were detected in all groups, but we inferred that Moraxella sp. are only significant in the early stages of the disease. M. conjunctivae was detected in all tested groups, while M. agalactiae was detected in samples of group III only. One strain of Moraxella sp. was resistant to erythromycin and showed intermedite resistance to tetracycline. Conclusion. The presence of these species confirms their importance in the aetiology of this disease, and the low resistance profile observed in the studied farm suggested an increased cure success rate.

Brazil has a herd of over 9 million goats, and the northeast of Brazil is home to over 93% of this herd. Caprine mycoplasmosis are widely disseminated worldwide, being highly contagious with high rates of morbidity and mortality, causing considerable economic loss to goat herders. In addition, there has been a lack of research using molecular testing to monitor the health and detect Mollicutes in this herd in Brazil. Therefore, the aim of this study is to associate animal management with the presence of the caprine origin Mollicutes in goats, in the southwest region of the state of Bahia, Brazil. A cross-sectional study was conducted on twelve farms, and statistical analyses were performed to identify associations between the presence of Mollicutes and the management of goats. Molecular testing identified Mollicutes class, Mycoplasma agalactiae (Ma) and M. conjunctivae (Mc), in the samples analyzed. Statistical associations were observed between animals from intensive livestock facilities and the presence of Mollicutes in nasal samples and dairy ranch animals and the presence of Mollicutes in ocular samples and animals from extensive ranching sites and positive results of Mollicutes in genital samples. We conclude that mycoplasmas are present in goat herds in the southwestern region of Bahia, which supports the need for more focused studies of mycoplasmas throughout the country. Our research also demonstrated the presence of two important opportunistic bacteria, Mc and Ma, and, to the best of our knowledge, this is the first time that M. conjunctivae was detected in Brazilian goats by molecular testing. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s42770-021-00517-z.

espanolLos micoplasmas son responsables de varias infecciones en pequenos rumiantes. Ya se han detectado micoplasmas potencialmente patogenos en el conducto auditivo externo de cabras y bovinos sanos brasilenos, y en asociacion con acaros, pero no en ovejas. La presente investigacion tuvo como objetivo evaluar el papel de la conjuntiva y el oido externo como fuente silenciosa de micoplasmas en ovejas. Se obtuvieron muestras de frotis del conducto auditivo externo y ocular de cuarenta y cinco ovejas machos y hembras. Deteccion molecular (PCR) y aislamiento de Mycoplasma spp. fueron realizados. Se determino la concentracion inhibitoria minima para el isolado utilizando oxitetraciclina, cloranfenicol, tilosina y acido nalidixico. Describimos el aislamiento de Mycoplasma conjunctivae a partir de la conjuntiva ocular de una oveja sana, ademas de la deteccion molecular de M. agalactiae y M. mycoides subsp. capri en ambos sitios anatomicos en ovinos del Estado de Sao Paulo, Brasil. El analisis filogenetico revelo proximidad y similitud con un aislado de oveja y de cabra estadounidenses. La conjuntiva ocular y el oido externo de las ovejas son criadores de micoplasmas veterinarios potencialmente patogenos que pueden infectar a las ovejas vulnerables. EnglishMycoplasmas are responsible for several infections in small ruminants. Potentially pathogenic mycoplasmas have al-ready been detected in the external ear canal of Brazilian healthy goats and cattle, and in association with mites, but not in sheep. The present research aimed to evaluate the role of the conjunc-tiva and external ear as a silent source of micoplasmas in sheep. Ocular and external ear canal swab samples were obtained from forty-five male and female sheep. Molecular detection (PCR) and isolation of Mycoplasma spp. were performed. Minimum inhibitory concentration was performed using oxytetracycline, tylosin, chloramphenicol and nalidixic acid. We describe the isolation of Mycoplasma conjunctivae fromthe ocular conjunc-tivae of a healthy sheep, besides the molecular detection of M. agalactiae and M. mycoides subsp. capri in both anatomical sites in sheep from the State of Sao Paulo. Phylogenetic anal-ysis revealed proximity and similarity with an American sheep and goat isolates. The ocular conjunctiva and the external ear of sheep are keepers of potentially pathogenic veterinary myco-plasmas which can infect vulnerable sheep

Although infectious keratoconjunctivitis (IKC) has occurred worldwide, no report has mentioned an IKC case in a dairy goat farm in an indoor management system. Moreover, few clinicians follow the IKC syndrome development throughout the clinical course. The aim of the present study was to report an IKC outbreak caused by Mycoplasma conjunctivae (MC) in indoor dairy goat barns. Severe IKC breakouts had been found consecutively in two nearby goat barns in a cooperated farm. To confirm the etiology, samples were collected from the infected goats to perform hematological, biochemical, bacterial examinations and polymerase chain reactions. A treatment of 4% boric acid and 0.5% povidone iodine solution, and a preventive spray of quaternary ammonium at 09:00 AM and 1% Virkon-S[Formula: see text] solution at 17:00 PM were useful to control disease in the adult goat barn, wherein 60% (31/55) animals showed different IKC signs. A total of 12 (14%) goat kids, died, without treatment, showed variable degrees of bronchointerstitial pneumonia. In all cases, only the DNA fragments of MC, but not other pathogens implicated in IKC, were detected. Goats began to recover after 30 days in an episodic duration of 81 days resulting in an estimated 240,000 New Taiwan dollars loss. In conclusion, this is the first goat MC report in an indoor system. The treatment and the preventive measure described in the current study were useful to control the event.

In late summer 2014, an outbreak of ocular disease occurred in the Norwegian muskox (Ovibos moschatus) population. Animals showed rings of pus around their eyes and one euthanized animal was diagnosed with acute keratoconjunctivitis. The DNA sequence analysis of eye-swab samples from this animal revealed a high abundance of Mycoplasma conjunctivae.

Resource competition and disease transmission may occur when domestic and wild ungulates live sympatricly. We investigated if the release of sheep (Ovis aries) onto alpine pasture in Switzerland affected chamois (Rupicapra rupicapra) activity budgets, local population size and spatial distribution. We also evaluated the risk of transmission of Mycoplasma conjunctivae (causing a contagious eye disease) from sheep to chamois by examining if the two species had close contact with one another. We carried out the study in an alpine valley containing two adjacent areas: one containing sheep (Fochsenflue) and one where sheep were excluded (Spitzflue). We found no difference between the activity budgets of the chamois at the two sites. At the Fochsenflue, chamois and sheep mainly used separate areas. However, after approximately 1month, sheep started to move twice per day, into the main area of the chamois. The percentage time feeding, spatial distribution and numbers of chamois did not change in response. Sheep were responsible for all encounters in which the two species came closer than 50m to each other. The encounters were brief, body contact never occurred, they were not concentrated at saltlicks and chamois mainly ended them. The results suggest that the presence of sheep had little effect on the chamois. However, competition between the two species could still be occurring over a longer time scale. Finally, we found that the risk of inter-specific transmission of IKC through direct body contact is likely to be low, but the risk through indirect means (flies or aerosols) remains

Infectious keratoconjunctivitis (IKC) caused by Mycoplasma conjunctivae is a widespread ocular affection of free-ranging Caprinae in the Alpine arc. Along with host and pathogen characteristics, it has been hypothesized that environmental factors such as UV light are involved in the onset and course of the disease. This study aimed at evaluating the role of topographic features as predisposing or aggravating factors for IKC in Alpine chamois (Rupicapra rupicapra rupicapra) and Alpine ibex (Capra ibex ibex). Geospatial analysis was performed to assess the effect of aspect (northness) and elevation on the severity of the disease as well as on the mycoplasmal load in the eyes of affected animals, using data from 723 ibex and chamois (583 healthy animals, 105 IKC-affected animals, and 35 asymptomatic carriers of M. conjunctivae), all sampled in the Swiss Alps between 2008 and 2010. An influence of northness was not found, except that ibex with moderate and severe signs of IKC seem to prefer more north-oriented slopes than individuals without corneal lesions, possibly hinting at a sunlight sensitivity consequent to the disease. In contrast, results suggest that elevation influences the disease course in both ibex and chamois, which could be due to altitude-associated environmental conditions such as UV radiation, cold, and dryness. The results of this study support the hypothesis that environmental factors may play a role in the pathogenesis of IKC.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://www.sciencedirect.com/science/article/pii/S0378113518305959?via%3Dihub This is the peer reviewed version of the following article: Aguilar, X.F., López-Olvera, J.R., Ribas, M.P., Begovoeva, M., Velarde, R., Cardells, J. et al. (2019). Mycoplasma conjunctivae in insect vectors and anatomic locations related to transmission and persistence. Veterinary Microbiology, vol. 228 (jan.), pp. 7-11, which has been published in final form at https://doi.org/10.1016/j.vetmic.2018.11.004. Este es el postprint del siguiente artículo: Aguilar, X.F., López-Olvera, J.R., Ribas, M.P., Begovoeva, M., Velarde, R., Cardells, J. et al. (2019). Mycoplasma conjunctivae in insect vectors and anatomic locations related to transmission and persistence. Veterinary Microbiology, vol. 228 (jan.), pp. 7-11, que se ha publicado de forma definitiva en https://doi.org/10.1016/j.vetmic.2018.11.004. Mycoplasma conjunctivae is an obligate microparasite that causes Infectious Keratoconjunctivitis (IKC) in Caprinae species. IKC is a long-recognised disease, but little attention has been paid to the mechanisms of transmission of the mycoplasma and its occurrence in locations other than the eyes. In this study, the presence of M. conjunctivae is assessed in the eyes, external ear canals (EEC), nasal cavity, and vagina of host species as well as in potential vectors, which may be involved in the transmission and persistence of infection within the host. M. conjunctivae was detected by qPCR in 7.2 % (CI 95% 4.7-11.0) of the ear swabs and 9.5 % (CI 95% 6.4-13.9) of the nasal swabs from Pyrenean chamois, Iberian ibex, domestic sheep and mouflon without statistical differences between species. Mycoplasma detection in nasal swabs was mostly associated with ocular infection (95.6%), but this was not the case for EEC (52.6%). Among the eye-positive ruminants, 27.3% were positive in ear swabs and 64.7% in nasal swabs, and the threshold cycle values of the qPCR were correlated only between eye and nasal swabs (p

Between 2006 and 2008, an outbreak of Infectious Keratoconjunctivitis (IKC) affected Pyrenean chamois Rupicapra p. pyrenaica, an endemic subspecies of mountain ungulate that lives in the Pyrenees. The study focused on 14 mountain massifs (180,000 ha) where the species’ population is stable. Cases of IKC were detected in ten of the massifs and, in five of them, mortality was substantial. The outbreak spread quickly from the first location detected, with two peaks in mortality that affected one (2007) and three (2008) massifs. In the latter, the peak was seasonal (spring to autumn) and, in the former, the outbreak persisted through winter. To identify the outbreak’s aetiology, we examined 105 Pyrenean chamois clinically affected with IKC. TaqMan rt-PCR identified Mycoplasma conjunctivae in 93 (88.5%) of the chamois. Another rt-PCR detected Chlamydophila spp. in 14 of chamois, and 12 of those had mixed infections with mycoplasmas. In the period 2000–2007, the chamois population increased slightly (λ 1.026) but decreased significantly during the IKC outbreak (λ 0.8, 2007–2008; λ 0.85, 2008–2009) before increasing significantly after the outbreak (λ 1.1, 2009–2010). Sex-biased mortality shifted the adult sex ratio toward males (from 0.6 to 0.7 males per female) and reduced productivity slightly. Hunting was practically banned in the massifs where chamois experienced significant mortality and allowed again after the outbreak ended. Long-term monitoring of wild populations provides a basis for understanding the impacts of disease outbreaks and improves management decisions, particularly when species are subject to extractive exploitation.

The susceptibility of the Iberian ibex ( Capra pyrenaica ) to Mycoplasma conjunctivae ocular infection and the changes in their interaction over time were studied in terms of clinical outcome, molecular detection, and IgG immune response in a captive population that underwent a severe infectious keratoconjunctivitis (IKC) outbreak. Mycoplasma conjunctivae was detected in the Iberian ibex, coinciding with the IKC outbreak. Its prevalence had a decreasing trend in 2013 that was consistent with the clinical resolution (August, 35.4%; September, 8.7%; November, 4.3%). Infections without clinical outcome were, however, still detected in the last handling in November. Sequencing and cluster analyses of the M. conjunctivae strains found 1 year later in the ibex population confirmed the persistence of the same strain lineage that caused the IKC outbreak but with a high prevalence (75.3%) of mostly asymptomatic infections and with lower DNA load of M. conjunctivae in the eyes (mean quantitative PCR [qPCR] cycle threshold [ C T ], 36.1 versus 20.3 in severe IKC). Significant age-related differences of M. conjunctivae prevalence were observed only under IKC epizootic conditions. No substantial effect of systemic IgG on M. conjunctivae DNA in the eye was evidenced with a linear mixed-models selection, which indicated that systemic IgG does not necessarily drive the resolution of M. conjunctivae infection and does not explain the epidemiological changes observed. The results show how both epidemiological scenarios, i.e., severe IKC outbreak and mostly asymptomatic infections, can consecutively occur by entailing mycoplasma persistence. IMPORTANCE Mycoplasma infections are reported in a wide range of epidemiological scenarios that involve severe disease to asymptomatic infections. This study allows a better understanding of the transition between two different Mycoplasma conjunctivae epidemiological scenarios described in wild host populations and highlights the ability of M. conjunctivae to adapt, persist, and establish diverse interactions with its hosts. The proportion of asymptomatic and clinical M. conjunctivae infections in a host population may not be regarded only in response to intrinsic host species traits (i.e., susceptibility) but also to a specific host-pathogen interaction, which in turn influences the infection dynamics. Both epidemic infectious keratoconjunctivitis and a high prevalence of asymptomatic M. conjunctivae infections may occur in the same host population, depending on the circulation of M. conjunctivae , its maintenance, and the progression of the host-pathogen interactions.

Functional roles of domestic and wild host populations in infectious keratoconjunctivitis (IKC) epidemiology have been extensively discussed claiming a domestic reservoir for the more susceptible wild hosts, however, based on limited data. With the aim to better assess IKC epidemiology in complex host-pathogen alpine systems, the long-term infectious dynamics and molecular epidemiology of Mycoplasma conjunctivae was investigated in all host populations from six study areas in the Pyrenees and one in the Cantabrian Mountains (Northern Spain). Detection of M. conjunctivae was performed by qPCR on 3600 eye swabs collected during seven years from hunted wild ungulates and sympatric domestic sheep (n = 1800 animals), and cluster analyses of the strains were performed including previous reported local strains. Mycoplasma conjunctivae was consistently detected in three Pyrenean chamois (Rupicapra p. pyrenaica) populations, as well as in sheep flocks (17.0% of sheep) and occasionally in mouflon (Ovis aries musimon) from the Pyrenees (22.2% in one year/area); statistically associated with ocular clinical signs only in chamois. Chamois populations showed different infection dynamics with low but steady prevalence (4.9%) and significant yearly fluctuations (0.0%- 40.0%). Persistence of specific M. conjunctivae strain clusters in wild host populations is demonstrated for six and nine years. Cross-species transmission between chamois and sheep and chamois and mouflon were also sporadically evidenced. Overall, independent M. conjunctivae sylvatic and domestic cycles occurred at the wildlife-livestock interface in the alpine ecosystems from the Pyrenees with sheep and chamois as the key host species for each cycle, and mouflon as a spill-over host. Host population characteristics and M. conjunctivae strains resulted in different epidemiological scenarios in chamois, ranging from the fading out of the mycoplasma to the epidemic and endemic long-term persistence. These findings highlight the capacity of M. conjunctivae to establish diverse interactions and persist in host populations, also with different transmission conditions.

Infectious keratoconjunctivitis (IKC) is a contagious eye disease primarily caused by Mycoplasma conjunctivae in domestic and wild Caprinae. Chlamydophila species have also been detected in ruminants with IKC. The objectives of this study are to investigate the ocular infection of M. conjunctivae and Chlamydiaceae and assess its interaction in relation to IKC in sheep and goats from remote communities around the Central Karakoram National Park in Pakistan, performing a combination of cross-sectional and case-control study design. Mostly asymptomatic and endemic infections of M. conjunctivae and Chlamydiaceae were found in sheep (19.3 per cent and 4.5 per cent, respectively) and goats (9.5 per cent and 1.9 per cent, respectively) from all communities, assessed by qPCR. Prevalence significantly differed between species only for M. conjunctivae (P=0.0184), which was also more prevalent in younger sheep (P

La queratoconjuntivitis infecciosa (QCI) és una malaltia ocular que afecta a ovins i caprins, tant domèstics com salvatges. La seva epidemiologia pot arribar a ser complexa i involucrar diverses espècies d’hostes a la interfase domèstic-salvatge de zones alpines. En aquest context, hipòtesis contràries sobre espècies reservori s’han formulat mitjançant dades epidemiològiques poc concloents. L’Objectiu principal d’aquesta tesi és proporcionar informació epidemiològica complerta per tal de inferir la funcionalitat de les diferents poblacions d’hostes presents als sistemes muntanyosos. En l’Estudi I i II, la situació epidemiològica de Mycoplasma conjunctivae es va avaluar en els ovins i caprins domèstics de tres zones muntanyoses, els Pirineus, la Serralada Cantàbrica (Nord Espanya) i la serralada del Karakorum (Himàlaia, Pakistan) per a tal d’avaluar quin risc poden suposar com a font de QCI per a la fauna salvatge simpàtrica. L’elevada prevalença trobada, junt amb la pràctica absència de signes clínics, suggereix que l’ovella (Ovis aries) és un bon hoste per al manteniment endèmic de M. conjunctivae. A més, Chlamydiaceae es va trobar de forma endèmica als ulls dels remugants domèstics de Pakistan i podria ser un factor predisposant per al desenvolupament clínica de QCI (Estudi II). A l’Estudi III, la dinàmica de M. conjunctivae i la seva epidemiologia molecular es va estudiar en les comunitats d’hostes domèstics i salvatges del Pirineu i la Serralada Cantàbrica. Els resultats van mostrar cicles epidemiològics selvàtics i domèstics separats al Pirineu. Clústers específics de soques de M. conjunctivae van ser mantinguts en algunes poblacions d’isards pirinencs (Rupicapra p. pyrenaica) sense la contribució substancial d’altres hostes. A més, es van observar diferents escenaris epidemiològics en les poblacions d’isard que van anar de la desaparició de M. conjunctivae a la persistència epidèmica i endèmica del micoplasma. La transmissió interespecífica de M. conjunctivae va ser puntualment constatada entre isard, mufló i ovella. Tot i que la gran majoria de casos de QCI trobats a l’isard no van ser originats per soques d’origen domèstic, les ovelles possiblement han tingut un paper històric important en la introducció de M. conjunctivae en les poblacions de fauna salvatge. Això és suggerit als Estudis II i III degut al consistent escenari epidemiològic trobat entre ovelles i isards simpàtrics. En l’Estudi IV, el curs d’una epizoòtia de QCI va ser estudiat en una població captiva de cabra salvatge (Capra pyrenaica), en la que la interacció micoplasma-hoste va evolucionar cap a signes clínics més lleus, menor càrrega bacteriana i possiblement una major persistència ocular del micoplasma. A més, aquest estudi proporciona evidències sobre el potencial rol que poden jugar els hostes salvatges en el manteniment de M. conjuntivae en diferents situacions epidemiològiques. Finalment, a l’Estudi V, es va explorar la presència de M. conjuntivae en localitzacions diferents als ulls i en relació amb la transmissió i persistència individual. La transmissió de M. conjuntivae és possible a través del contacte directe amb secrecions oculars i nasals i de forma indirecta mitjançant vectors (Musca sp.). A més, Mycoplasma conjunctivae es va detectar al conducte auditiu de diferents espècies d’hostes, fet que pot tenir implicacions epidemiològiques importants. En conclusió, els resultats d’aquesta tesi ressalten el capacitat de M. conjuntivae per a establir diferents interaccions amb els seus hostes i persistir a les poblacions mitjançant patrons de transmissió diferents. El paper de reservori o d’hospedador accidental està determinat tant per factors relacions amb el propi hospedador com amb el micoplasma. Les característiques de les poblacions d’hostes possiblement també modulen la interacció micoplasma-hoste mitjançant la connectivitat entre individus, la transmissió de M. conjuntivae i finalment determinar la seva persistència i el paper dels diferents hostes en el sistema., La queratoconjuntivitis infecciosa (QCI) es una enfermedad ocular que afecta a ovinos y caprinos, tanto domésticos como salvajes. Su epidemiología puede ser compleja e involucrar diversas especies de hospedadores en la interfaz domestico-salvaje de zonas alpinas. En este contexto, hipótesis contrarias sobre especies reservorio se han formulado mediante datos epidemiológicos poco concluyentes. El objetivo principal de esta tesis es proporcionar información epidemiológica completa a fin de inferir la funcionalidad de los distintos hospedadores presentes en los sistemas montañosos. En el Estudio I y II, la situación epidemiológica sobre Mycoplasma conjunctivae se evaluó en ovinos y caprinos domésticos de tres zonas montañosas, Pirineo y Cordillera cantábrica (Norte Español) y la Cordillera del Karakorum (Himalaya, Pakistán), a fin de evaluar el riesgo que éstos pueden suponer como fuente de QCI para la fauna. Mycoplasma conjunctivae fue detectado frecuentemente en los rebaños de ovejas y cabras que pastan en las zonas alpinas de los Pirineos y del Karakorum, y de forma más esporádica en la Cordillera Cantábrica. La alta prevalencia encontrada, junto con la práctica ausencia de enfermedad ocular, sugiere que la oveja doméstica (Ovis aries) es un buen hospedador para el mantenimiento endémico de M. conjunctivae. Además, Chlamydiaceae se encontró presente en los ojos de rumiantes de Pakistán y podría ser un factor predisponente para el desarrollo clínico de la QCI (Estudio II). En el Estudio III, la dinámica de M. conjunctivae y su epidemiología molecular fue estudiada en la comunidad de hospedadores domésticos y salvajes del Pirineo y la Cordillera Cantábrica. Los resultados mostraron ciclos epidemiológicos selváticos y domésticos separados. Se observaron distintos escenarios epidemiológicos en las poblaciones de rebeco (Rupicapra p. pyrenaica), que fueron de la desaparición de M. conjunctivae a la persistencia endémica y epizoótica del micoplasma. La transmisión interespecífica de M. conjunctivae fue puntualmente constatada entre rebeco, muflón y oveja. A pesar de que la mayoría de casos de QCI encontrados en le rebeco no fueron originados por cepas de rebaños domésticos, las ovejas posiblemente han tenido un papel histórico importante en la introducción de M. conjunctivae en las poblaciones de fauna salvaje. Esto es sugerido en los Estudio II y III debido al consistente escenario epidemiológico encontrado entre ovejas y rebecos simpátricos. En el Estudio IV, el curso de una epizootía de QCI fue estudiada a largo plazo en una población cautiva de cabra montés (Capra pyrenaica), en la que la interacción micoplasma-hospedador evolucionó hacia signos clínicos más leves, menor carga bacteriana y posiblemente una mayor persistencia ocular del micoplasma. Este estudio además proporciona evidencias sobre el potencial rol que pueden jugar los hospedadores salvajes en el mantenimiento de M. conjunctivae en relación a distintas situaciones epidemiológicas. Finalmente, en el Estudio V, se exploró la presencia de M. conjunctivae en localizaciones distintas a los ojos en relación con su transmisión y persistencia individual. La transmisión de M. conjunctivae es posible a través de contacto directo con secreciones oculares y nasales y de forma indirecta mediante vectores (Musca sp.). Mycoplasma conjunctivae además se detectó en el conducto auditivo, lo cual puede tener implicaciones epidemiológicas importantes. En conclusión, los resultados de esta tesis resaltan la capacidad de M. conjunctivae para establecer distintas interacciones con sus hospedadores y persistir en las poblaciones mediante patrones de transmisión distintos. El papel de reservorio o hospedador accidental está determinado tanto por factores relacionados con el propio hospedador como con el micoplasma. Las características de las poblaciones susceptibles posiblemente también modulan la interacción micoplasma-hospedador mediante la conectividad entre individuos, transmisión de M. conjunctivae y finalmente determinar su persistencia y el papel de los distintos hospedadores en el sistema., Infectious keratoconjunctivitis (IKC) is an ocular disease that affects wild and domestic Caprinae species. Epidemiology of IKC may rely in complex multi-host systems at the wildlife-livestock interface of alpine areas, where contrasting hypotheses of reservoir hosts have been proposed based on limited data. The main objective of this thesis is to provide comprehensive epidemiological data in order to infer the functional roles of domestic and wild host populations from mountain systems. In the Study I and Study II, the epidemiological situation of Mycoplasma conjunctivae in domestic small ruminants from mountain areas in two distant regions (Northern Spain and Pakistan), was studied in order to assess the potential source risk they suppose for their sympatric wild counterparts. Mycoplasma conjunctivae was commonly detected in the Pyrenees and the Karakoram mountain range, and more rarely in the Cantabrian Mountains. The relatively high prevalence of mostly asymptomatic infections observed suggests that sheep (Ovis aries) is a good host for the endemic maintenance of M. conjunctivae with a low cost (i.e. clinical signs and body condition). Chlamydiaceae was also endemic in the eyes of sheep and goats (Capra hircus) from Pakistan and could be a predisposing factor for IKC that needs of further research (Study II). The long-term dynamics of M. conjunctivae and its molecular epidemiology in the whole community of wild and domestic hosts from the Pyrenees and Cantabrian Mountains was assessed in the Study III. Results showed that independent M. conjunctivae sylvatic and domestic cycles occurred at the wildlife-livestock interface in the Pyrenees. Specific M. conjunctivae strain clusters were maintained in some Pyrenean chamois (Rupicapra p. pyrenaica) populations without the substantial contribution of other hosts. Furthermore, host population characteristics and M. conjunctivae strains resulted in different epidemiological scenarios in chamois, ranging from the fading out and the epidemic persistence in lower density populations to the endemic persistence in a higher density population. Nevertheless, incidental cross-species transmission of M. conjunctivae occurred between chamois and mouflon and between chamois and sheep. Despite most of the IKC cases in free-ranging chamois were not associated with concurrent sheep strains, domestic sheep may have been historically important for the introduction of M. conjunctivae in wild hosts. This is suggested in the Studies II and III by the consistent epidemiological scenario observed in sympatric sheep and chamois from the Cantabrian Mountains and the Pyrenees. In the Study IV, the transition from epizootic IKC to high prevalence of asymptomatic M. conjunctivae infections is described in a captive Iberian ibex (Capra pyrenaica) population, in which the host-mycoplasma interaction evolved to decreased clinical signs, lower mycoplasma loads and probably longer persistence in the eyes (Study IV). This study also provides evidences of the potential role of wild hosts in M. conjunctivae maintenance, associated to different and dynamic epidemiological scenarios. Finally, in the Study V, the detection of M. conjunctivae in other locations apart from the eyes was explored in different host species in relation to transmission routes and within-host persistence. Transmission of Mycoplasma conjunctivae has been reasserted to be possible through direct contact from ocular and nose secretions and indirectly by eye-frequenting flies (Musca sp.). Mycoplasma conjunctivae was also detected in the ear canals, which may have important epidemiological implications. Overall, the results obtained highlight the capacity of M. conjunctivae to establish diverse interactions with its hosts and persist in the population, also with different transmission patterns. Host populations and species can act either as maintenance hosts or as spill-over hosts depending on host- and M. conjunctivae-related factors. Population characteristics may also shape host-mycoplasma interaction through connectivity among individuals, M. conjunctivae transmission, and ultimately may determine its persistence and host functionalities in the system.

Clinicians can evaluate the relevance of an outbreak based on its basic reproductive number (R0). So far there has been no report on the R0 of Mycoplasma conjunctivae which is a major cause of goats' conjunctivitis in Taiwan. The present study sought to investigate an outbreak of infectious keratoconjunctivitis (IKC) by Mycoplasma conjunctivae (MC) in an indoor dairy goat barn. The epidemiological curve was recorded to build a susceptible-infected-recovered model and to estimate the R0 by three methods In the investigated goat barn, 60% (31/55) goats showed degrees of IKC signs. The number of infected animals increased quickly after 15days, but slowed down after 41days. The sick goats began to recover after 30days. The epidemic fully stopped after 81days. The estimated R0 ranged from 1.35 to 4.46. In summary, this is the first MC report in Taiwan, and the first one to estimate the R0 of MC.

Mycoplasma conjunctivae are etiological agents of infectious keratoconjunctivitis (IKC), commonly known as pink-eye in domestic sheep, goats and other wild animals in many parts of the world. A few young Lohi lambs maintained at Livestock Experiment Station (LES), Bahadurnagar, Okara, Pakistan showed clinical signs and symptoms of conjunctivitis, keratitis, severe lacrimation and varying degree of blindness. During January to March, 2011, a total of 36 ocular swabs were collected from IKC affected animals and were processed for isolation, identification, and characterization of M. conjunctivae. Sixteen (44.44 %) out of 36 samples showed turbidity in PPLO broth. Twelve (75 %) out of 16 broth samples showed colony growth on PPLO agar. All 16 (44.44 %) out of 36 turbid broth samples, 12 (75 %) out of 16 cultured on agar plate samples, and 21 (59 %) out of 36 sheep ocular direct swab samples were found positive for M. conjunctivae through polymerase chain reaction test by using M. conjunctivae-specific primer pair McoF1 and McoR1 and detecting a 750 base pair fragment on agarose gel. Topical application of 0.5 % sterile solution of gentamycin (100 mg/ml) (Gentafar 10 %, FARVET, Netherlands) proved suitable for the treatment of IKC in Lohi lambs as all clinical signs of IKC disappeared after 5 days of treatment with this antibiotic. This is the first report about the prevalence, molecular diagnosis, and treatment of M. conjunctivae in Lohi sheep affected with infectious keratoconjunctivitis at LES, Bahadurnagar, Okara, Pakistan.

Background Infectious keratoconjunctivitis (IKC) is an ocular infectious disease caused by Mycoplasma conjunctivae which affects small domestic and wild mountain ruminants. Domestic sheep maintain the pathogen but the detection of healthy carriers in wildlife has raised the question as to whether M. conjunctivae may also persist in the wild. Furthermore, the factors shaping the dynamics of IKC outbreaks in wildlife have remained largely unknown. The aims of this study were (1) to verify the etiological role of M. conjunctivae in IKC outbreaks recorded between 2002 and 2010 at four study sites in different regions of France (Pyrenees and Alps, samples from 159 Alpine ibex Capra ibex, Alpine chamois Rupicapra rupicapra and Pyrenean chamois Rupicapra pyrenaica); (2) to establish whether there existed any epidemiological links between the different regions through a cluster analysis of the detected strains (from 80 out of the 159 animals tested); (3) to explore selected pathogen, host and environmental factors potentially influencing the dynamics of IKC in wildlife, by joining results obtained by molecular analyses and by field observations (16,609 animal observations). All of the samples were tested for M. conjunctivae by qPCR, and cluster analysis was based on a highly variable part of the lppS gene. Results We documented infections with M. conjunctivae in epidemic and endemic situations, both in symptomatic and asymptomatic animals. The identified M. conjunctivae strains were site-specific and persisted in the local wild population for at least 6 years. In epidemic situations, peaks of cases and disease resurgence were associated with the emergence of new similar strains in a given area. Social interactions, seasonal movements and the landscape structure such as natural and anthropogenic barriers influenced the spatio-temporal spread of IKC. Adults were more affected than young animals and host susceptibility differed depending on the involved strain. Conclusion Our study indicates that IKC is a multifactorial disease and that M. conjunctivae can persist in wildlife populations. The disease course in individual animals and populations is influenced by both host and mycoplasma characteristics, and the disease spread within and among populations is shaped by host behavior and landscape structure. Electronic supplementary material The online version of this article (doi:10.1186/s12917-017-0972-0) contains supplementary material, which is available to authorized users.

Mycoplasma conjunctivae, the causative agent of infectious keratoconjunctivitis (IKC), was recently detected in asymptomatic Alpine ibex (Capra ibex ibex). This suggested that an external source of infection may not be required for an IKC outbreak in wildlife but might be initiated by healthy carriers, which contradicted previous serologic investigations in chamois. Our aims were to 1) assess the prevalence of M. conjunctivae among asymptomatic ibex and Alpine chamois (Rupicapra rupicapra rupicapra) and its frequency in IKC-affected animals, 2) determine mycoplasma loads in different disease stages, and 3) characterize the M. conjunctivae strains involved. Eye swabs from 654 asymptomatic and 204 symptomatic animals were collected in diverse Swiss regions between 2008 and 2010, and tested by TaqMan real-time PCR. Data analysis was performed considering various patterns of IKC occurrence in the respective sampling regions. Strains from 24 animals were compared by cluster analysis. Prevalence of M. conjunctivae was 5.6% (95% confidence interval [CI]: 3.7-8.1%) in asymptomatic ibex and 5.8% (CI: 3.0-9.9%) in asymptomatic chamois, with significant differences between years and regions in both species. Detection frequency in symptomatic animals was significantly higher during IKC outbreaks than in nonepidemic situations (i.e., regular but low incidence or sporadic occurrence). Mycoplasma load was significantly lower in eyes from healthy carriers and animals with mild signs than from animals with moderate and severe signs. Although some strains were found in both asymptomatic and diseased animals of the same species, others apparently differed in their pathogenic potential depending on the infected species. Overall, we found a widespread occurrence of M. conjunctivae in wild Caprinae with and without IKC signs. Our results confirm the central role of M. conjunctivae in outbreaks but suggest that other infectious agents may be involved in IKC cases in nonepidemic situations. Additionally, presence and severity of signs are related to the quantity of M. conjunctivae in the eyes rather than to the strain. We propose that individual or environmental factors influence the clinical expression of the disease and that persistence of M. conjunctivae in populations of wild Caprinae cannot be excluded.

Because interactions between livestock and chamois occur on Alpine pastures, transmission of infectious diseases is considered possible. Thus, the occurrence of Chlamydiaceae, Mycoplasma conjunctivae, and pestiviruses in Alpine chamois ( Rupicapra r. rupicapra) of the Surselva region (eastern Swiss Alps) was investigated. In total, 71 sera, 158 eye swabs, 135 tissue samples, and 23 fecal samples from 85 chamois were analyzed. The sera were tested by 2 enzyme-linked immunosorbent assay (ELISA) kits specific for Chlamydophila abortus. Eye swabs, tissue, and fecal samples were examined by a Chlamydiaceae-specific realtime polymerase chain reaction (PCR). Positive cases were further investigated by microarray method. One serum sample (1.4%) was positive in 1 of the ELISAs. Eye swabs of 3 chamois (3.8%) were positive for Chlamydiaceae. The microarray method revealed the presence of Chlamydophila abortus, C. pecorum, and C. pneumoniae. All tissue and fecal samples were negative. With real-time PCR, 3.9% of the chamois tested positive for Mycoplasma conjunctivae. One chamois had a simultaneous infection with M. conjunctivae and 2 chlamydial species ( C abortus, C. pecorum). Skin and tongue tissue samples of 35 chamois were negative for pestivirus antigen by immunohistochemistry. It was concluded that in contrast to the findings in Pyrenean chamois ( Capra p. pyrenaica) of Spain, the occurrence of Chlamydiaceae in Alpine chamois of the Surselva region is low, and the transmission between domestic and wild Caprinae seems not to be frequent. Comparably, persistent pestiviral infections do not seem to be common in chamois of the Surselva region.

Summary The bacteriological findings in infectious kerato-conjunctivitis of chamois and sheep in Switzerland are described. In addition to a relatively heterogeneous flora (Staph. aureus, Corynebacterium pyogenes, Moraxella ovis, Haemophilus sp., Actinobacillus lignieresii, micrococci and streptococci) we isolated constantly M. conjunctivae, sometimes in mixed infection with M. arginini. In one instance M. conjunctivae was associated with pneumonia. The possible role of M. conjunctivae as an etiological agent in infectious kerato-conjunctivitis is discussed. Zusammenfassung Nachweis von Mycoplasma conjunctivae bei der Keratoconjunctivitis der Gemsen und Schafe Bei der infektiosen Keratoconjunctivitis der Gemsen und Schafe konnte neben einer heterogenen Bakterienflora (Staph. aureus, Corynebacterium pyogenes, Moraxella ovis, Haemophilus sp., Actinobacillus lignieresii, Mikrokokken und Streptokokken) regelmasig M. conjunctivae isoliert werden. Zum Teil lag eine Mischinfektion mit M. arginini vor. In einem weiteren Fall trat M. conjunctivae im Zusammenhang mit einer Pneumonie auf. Die Befunde sprechen fur die aetiologische Bedeutung von M. conjunctivae fur die infektiose Keratoconjunctivitis. Resume Mise en evidence de Mycoplasma conjunctivae dans la Keratoconjonctivite des chamois et des moutons M. conjunctivae a ete regulierement isole dans la Keratoconjonctivite infectieuse des chamois et des moutons a cote d'une flore bacterienne heterogene (Staph. aureus, Corynebacterium pyogenes, Moraxella ovis, Haemophilus sp., Actinobacillus lignieresii, des microcoques et des streptocoques). On a parfois rencontre une infection mixte avec M. arginini. M. conjunctivae est intervenu dans d'autres cas en relation avec une pneumonie. Les resultats parlent en faveur du role etiologique de M. conjunctivae dans la Keratoconjonctivite infectieuse. Resumen Deteccion de Mycoplasma conjunctivae en la queratoconjuntivitis de las gamuzas y ovejas En la queratoconjuntivitis infecciosa de las gamuzas y ovejas se pudo aislar con regularidad Mycoplasma conjunctivae al lado de una flora bacteriana heterogenea (Staph. aureus, Corynebacterium pyogenes, Moraxella ovis, Haemophilus spp., Actinobacillus lignieresii, micrococos y estreptococos). En parte se trataba de una infeccion mixta con M. arginini. En otro caso mas se presento en relacion con una pneumonia. Los hallazgos abogan a favor de la importancia etiologica de M. conjunctivae para la queratoconjuntivitis infecciosa.

Infectious keratoconjunctivitis (IKC) is worldwide known as a common infection by Mycoplasma conjunctivae affecting the eyes of domestic sheep and goats where it generally leads to moderate symptoms known as ‘pink eye’. IKC also occurs in severe outbreaks in free-ranging alpine chamois Rupicapra r. rupicapra and alpine ibex Capra i. ibex. It is characterised by grave clinical symptoms resulting in blindness and perforations of the cornea of the affected animals, and finally leads to their death. Transmission of M. conjunctivae from domestic sheep to wild Caprinae has been demonstrated by molecular epidemiological studies of the infectious agent. The relatively high prevalence of M. conjunctivae in sheep populations that share pastures in the Alps with wild Caprinae, in which outbreaks occur infrequently but with high virulence, raised an important question concerning the role of host-specificity of various strains of M. conjunctivae. Investigations on genetic variations of M. conjunctivae strains...

The humoral immune response of three alpine chamois (Rupicapra rupicapra rupicapra), two alpine ibex (Capra ibex ibex) and three domestic sheep naturally affected with infectious keratoconjunctivitis (IKC), and four ibex and two sheep experimentally infected with Mycoplasma conjunctivae was analysed. In addition, the local immune response to M. conjunctivae was analysed using conjunctival washes from chamois and sheep. Immunoblot analysis of sera using whole cell antigens of M. conjunctivae revealed the major immunogenic proteins which had molecular masses of 175, 83, 68, 60, 50, 42, 36, and 33 kDa. Major antigens were found at 83, 68, 60, and 42 kDa in both sera and conjunctival washes from naturally infected animals of all three Caprinae species. In experimentally infected animals, antibodies to the 68 and 60 kDa antigens were dominant. Naturally infected animals showed much stronger immune reactions than those experimentally infected, and specific antibodies appeared 2 to 4 wk after experimental infection. To evaluate possible cross-reactions, whole cell antigen of M. conjunctivae was analysed by immunoblot against hyperimmune sera of closely related Mycoplasma spp. Antibodies to the 175, 73, 68, 60, and 33 kDa antigens appeared to be specific to M. conjunctivae. Cross-reactions mainly with 83, 50, and 42 kDa antigens were detected, in particular with M. ovipneumoniae and M. bovoculi hyperimmune sera, but also with antisera against M. capricolum capricolum and M. putrefaciens.

Background Infectious keratoconjunctivitis (IKC) is a clinical condition affecting eyes of domestic and wild Caprinae worldwide, and Mycoplasma conjunctivae is considered the primary causative agent of IKC in sheep, goats and wild Caprinae. Domestic ruminants from high mountain habitats share grazing areas with wild mountain ungulates, such as chamois (Rupicapra spp.), Alpine ibex (Capra ibex) and European mouflon (Ovis aries musimon), and domestic sheep seem to act as M. conjunctivae reservoir. In this study, the presence of M. conjunctivae in domestic sheep and goats from the two main mountain ranges of Northern Spain, the Pyrenees and the Cantabrian Mountains, has been investigated. Results Eye swabs were obtained from 439 domestic small ruminants selected from flocks that seasonally graze in alpine meadows during three consecutive years (2011-2012-2013). Seventy-nine out of the 378 domestic sheep (20.9%) tested positive to a M. conjunctivae specific real time-PCR (rt-PCR) in at least one eye, whereas all the 61 sampled domestic goats were negative. Statistically significant higher prevalence and higher proportion of infected flocks (P M. conjunctivae is widespread and probably endemic in domestic sheep, than in the Cantabrian Mountains (7.8%; one flock out of six). Twenty-five sheep (three from the Pyrenees and 22 from the Cantabrian Mountains) which showed clinical signs consistent with infectious keratoconjunctivitis (IKC) were negative by rt-PCR. In contrast, 62 out of the 71 (87.3%) M. conjunctivae-positive sheep from the Pyrenees and the eight positive sheep from the Cantabrian Mountains were asymptomatic. Conclusions This study provides rt-PCR-based evidences of M. conjunctivae maintenance in domestic sheep, as well as a relationship between prevalence in domestic sheep and previously reported M. conjunctivae and IKC in wild ruminants. Domestic goats do not seem to play an important role in the epidemiology of M. conjunctivae in alpine habitats from Northern Spain.

Summary Mycoplasma conjunctivae was isolated four times from the eyes of nine Alpine ibex (Capra ibex ibex) suffering from keratoconjunctivitis. The animals examined were affected in two different outbreaks in the Swiss Alps. Parasitological and bacteriological studies, including investigations for chlamydia and mycoplasmas, were performed. The results indicate that M. conjunctivae is the primary pathogenic agent causing infectious keratoconjunctivitis in this species.

The extent to which pathogens maintain the extraordinary polymorphism at vertebrate Major Histocompatibility Complex (MHC) genes via balancing selection has intrigued evolutionary biologists for over half a century, but direct tests remain challenging. Here we examine whether a well-characterized epidemic of Mycoplasmal conjunctivitis resulted in balancing selection on class II MHC in a wild songbird host, the house finch (Carpodacus mexicanus). First, we confirmed the potential for pathogen-mediated balancing selection by experimentally demonstrating that house finches with intermediate to high multi-locus MHC diversity are more resistant to challenge with Mycoplasma gallisepticum. Second, we documented sequence and diversity-based signatures of pathogen-mediated balancing selection at class II MHC in exposed host populations that were absent in unexposed, control populations across an equivalent time period. Multi-locus MHC diversity significantly increased in exposed host populations following the epidemic despite initial compromised diversity levels from a recent introduction bottleneck in the exposed host range. We did not observe equivalent changes in allelic diversity or heterozygosity across eight neutral microsatellite loci, suggesting that the observations reflect selection rather than neutral demographic processes. Our results indicate that a virulent pathogen can exert sufficient balancing selection on class II MHC to rescue compromised levels of genetic variation for host resistance in a recently bottlenecked population. These results provide evidence for Haldane's long-standing hypothesis that pathogens directly contribute to the maintenance of the tremendous levels of genetic variation detected in natural populations of vertebrates. Published version

SUMMARYThe conjunctival sacs of 100 healthy adult ewes and 20 lambs and 76 adult ewes affected by ovine keratoconjunctivitis (OKC) were examined microbiologically.Branhamella oviswas the only organism regularly isolated from healthy ewes and lambs.Mycoplasma conjunctivaewas isolated from the conjunctival sac of nine ewes but its occurrence was not associated with clinical signs of ovine keratoconjunctivitis.Mycoplasma conjunctivaeandBranhamella oviswere isolated significantly more often from eyes affected by OKC than from unaffected eyes.

Monitoraggio a lungo termine del camoscio dei Pirenei in una zona protetta: fluttuazione della popolazione. Tra il 1993 ed il 2009, la popolazione di camoscio pirenaico Rupicapra p. pyrenaica del Parco Nazionale d’Aigüestortes e Estany de Sant Maurici (14119 ha, Catalogna, Spagna) è stata monitorata periodicamente. Nel periodo post-riproduttivo (luglio), i censimenti sono stati condotti lungo 20 percorsi supraforestali fissi, con l’obiettivo di stimare le variazioni nelle dimensioni, rapporto sessi, struttura e produttività della popolazione nell’area protetta. La densità media è risultata pari a 6,7 camosci km-2, il rapporto sessi circa 1:3 in favore delle femmine, mentre la produttività è stata pari a 0,6 juvenes per femmina adulta. La proporzione di animali indeterminati durante i conteggi è diminuita significativamente nel tempo, probabilmente in seguito al progressivo training degli operatori. Nel periodo di studio, le dimensioni della popolazione sono fluttuate significativamente. Sebbene le cause di tali variazioni siano ancora da approfondire, le successive epidemie di cheratocongiuntivite infettiva e Pestivirus che hanno colpito la popolazione hanno probabilmente svolto un ruolo determinante. Si raccomanda di sviluppare un piano di gestione dell’intero massiccio montuoso, che includa regolari monitoraggi sanitari e conteggi autunnali.

Summary Ovine keratoconjunctivitis (OKC) identical to that seen in field cases was produced experimentally with 106 colony forming units of an isolate of Mycoplasma conjunctivae which had been passaged three times in vitro. Cytological studies showed that the peak neutrophil count in conjunctival exudate occurred 8 days after inoculation of the conjunctival sac. Rapid onset of clinical signs in the uninoculated eye showed the ease with which the organism could be transferred from affected to unaffected eyes.