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Topic:Infectious Disease

Infectious diseases in horses encompass a range of illnesses caused by bacteria, viruses, fungi, or parasites. These diseases can affect various systems within the equine body, leading to symptoms that range from mild discomfort to severe systemic illness. Common infectious diseases in horses include equine influenza, strangles, equine herpesvirus, and West Nile virus. These diseases can be transmitted through direct contact with infected animals, contaminated surfaces, or vectors such as insects. Understanding the mechanisms of transmission, pathogenesis, and immune response is essential for effective prevention and control. This page compiles peer-reviewed research studies and scholarly articles that explore the epidemiology, diagnosis, treatment, and management of infectious diseases in horses.
Clostridioides difficile infection in thoroughbred horses in Japan from 2010 to 2021.
Scientific reports    August 11, 2023   Volume 13, Issue 1 13099 doi: 10.1038/s41598-023-40157-x
Uchida-Fujii E, Niwa H, Senoh M, Kato H, Kinoshita Y, Mita H, Ueno T.We encountered 34 Clostridioides difficile (C. difficile) infection (CDI) cases among Thoroughbred horses in Japan from 2010 to 2021. Among them, 79.4% (27/34) either died or were euthanised. The risk factors associated with CDI and mortality among Japanese Thoroughbred horses remain unclear. We used genetic methods to examine C. difficile strains and their relationships with prognosis. Twenty-two (64.7%) cases were hospitalised at the onset of colitis. Outcomes were balanced for hospitalisation rates at the onset of colitis. The mortality rates of cases treated with metronidazole (65.0%) were...
Host-mosquito interactions in rural and urban equestrian facilities from temperate Argentina.
Medical and veterinary entomology    August 10, 2023   Volume 37, Issue 4 816-825 doi: 10.1111/mve.12686
Cardo MV, Vezzani D.Targeted vector surveillance informed by data on mosquito biting patterns can help limit arboviral zoonotic diseases. To characterise host-biting networks in rural and urban equestrian facilities from temperate Argentina, adult resting mosquitoes were collected (December 2018-April 2019) with a battery-powered aspirator. Engorged females were sorted to species, and their blood source was identified using molecular techniques. Bipartite network analysis was performed for rural and urban matrices. A total of 177 bloodmeals from 11 mosquito species of Aedes and Culex were identified, with seven m...
Distribution of West Nile virus cases in horses reveals different spatiotemporal patterns in eastern and western Canada.
Journal of the American Veterinary Medical Association    August 9, 2023   1-9 doi: 10.2460/javma.23.05.0259
Levasseur A, Arsenault J, Paré J.West Nile virus (WNV) became notifiable in horses in 2003 in Canada and has been reported every year since. The objective of this study was to describe the spatiotemporal distribution of WNV in horses between 2003 and 2020 in Canada. Methods: The 848 symptomatic and laboratory-confirmed WNV cases in horses reported to the Canadian Food Inspection Agency between 2003 and 2020. Methods: Canada was divided into eastern and western regions for analysis. For each case, location and date of notification were captured. Triennial maps were made to describe the spatiotemporal distribution and expansion...
A case of bacteremia and pneumonia caused by Streptococcus equi subspecies equi infection in a 70-year-old female following horse exposure in rural Wyoming.
Annals of clinical microbiology and antimicrobials    August 2, 2023   Volume 22, Issue 1 65 doi: 10.1186/s12941-023-00602-1
Bohlman T, Waddell H, Schumaker B.The occurrence of zoonotic infections following an animal exposure continues to be an important consideration for all patients, especially those within agricultural communities. Streptococcus equi subspecies equi (S. equi subsp. equi) is a bacteria known to cause a common infection called 'Strangles' in horses. This article highlights a new case of pneumonia and bacteremia in a patient caused by S. equi subsp. equi following strangles exposure in a horse. Rarely has there been reported horse to human transmission of subsp. equi. Methods: A 70-year-old woman attended a rural emergency departmen...
Validation of a serum ELISA test for cyathostomin infection in equines.
International journal for parasitology    August 1, 2023   doi: 10.1016/j.ijpara.2023.07.001
Lightbody KL, Austin A, Lambert PA, von Samson-Himmelstjerna G, Jürgenschellert L, Krücken J, Nielsen MK, Sallé G, Reigner F, Donnelly CG....Cyathostomins are ubiquitous equine nematodes. Infection can result in larval cyathostominosis due to mass larval emergence. Although faecal egg count (FEC) tests provide estimates of egg shedding, these correlate poorly with burden and provide no information on mucosal/luminal larvae. Previous studies describe a serum IgG(T)-based ELISA (CT3) that exhibits utility for detection of mucosal/luminal cyathostomins. Here, this ELISA is optimised/validated for commercial application using sera from horses for which burden data were available. Optimisation included addition of total IgG-based calibr...
Utility of serum amyloid A in monitoring clinical response to antimicrobial treatment in horses with bacterial pneumonia.
Journal of veterinary internal medicine    July 31, 2023   doi: 10.1111/jvim.16818
Hepworth-Warren KL, Estell K, Cowles B, Amodie D, Crisman M.Serum amyloid A (SAA) is a major acute phase protein in horses which could be a useful tool for assessing clinical response to treatment of bacterial pneumonia in adult horses.
Relationship between the Cycle Threshold Value (Ct) of a Salmonella spp. qPCR Performed on Feces and Clinical Signs and Outcome in Horses.
Microorganisms    July 30, 2023   Volume 11, Issue 8 1950 doi: 10.3390/microorganisms11081950
Amory H, Cesarini C, De Maré L, Loublier C, Moula N, Detilleux J, Saulmont M, Garigliany MM, Lecoq L.The objective of this retrospective study was to evaluate the clinical significance of fecal quantitative real-time polymerase chain reaction (qPCR) results when taking the cycle threshold values (Ct) into account. The study included 120 qPCR-positive fecal samples obtained from 88 hospitalized horses over a 2-year period. The mean Ct of the qPCR test was evaluated in regard to (1) clinical outcome and (2) systemic inflammatory response syndrome (SIRS) status (no SIRS, moderate SIRS, or severe SIRS) of the sampled horses. An ROC analysis was performed to establish the optimal cut-off Ct valu...
Transfer of naturally acquired specific passive immunity against Anaplasma phagocytophilum in foals in Southeastern Pennsylvania and Northern Maryland.
Journal of veterinary internal medicine    July 28, 2023   Volume 37, Issue 5 1889-1892 doi: 10.1111/jvim.16812
Rule EK, Boyle AG, Stefanovski D, Anis E, Linton J, Lorello O.Equine granulocytic anaplasmosis (EGA) is a common disease in adult horses, but clinical disease in foals is rarely reported. The relationship between equine maternal and neonatal antibodies to Anaplasma phagocytophilum is unclear. Objective: That mares in an endemic region would be seropositive for A. phagocytophilum and that mare and foal serum IgG concentrations for A. phagocytophilum would correlate. Additionally, we hypothesized that foal IgG concentrations for A. phagocytophilum acquired by passive immunity would decline by 6 months of age. Methods: Twenty-two healthy mare-foal pairs. ...
Equine Psittacosis and the Emergence of Chlamydia psittaci as an Equine Abortigenic Pathogen in Southeastern Australia: A Retrospective Data Analysis.
Animals : an open access journal from MDPI    July 28, 2023   Volume 13, Issue 15 2443 doi: 10.3390/ani13152443
El-Hage C, Legione A, Devlin J, Hughes K, Jenkins C, Gilkerson J. is an important zoonotic pathogen. Although primarily a pathogen of birds, from which infection can spillover into humans and other mammalian hosts, the importance of as a cause of equine reproductive loss and the risk of infection to humans in contact with infected horses are increasingly being recognised in Australia and elsewhere. Despite the risks to both human and equine health, infection in horses is incompletely understood. This study aimed to update and summarise cases of equine psittacosis in Australia in the period 2018-2022, thus addressing a knowledge gap relating to recent case...
Quadruplex Real-Time TaqMan® RT-qPCR Assay for Differentiation of Equine Group A and B Rotaviruses and Identification of Group A G3 and G14 Genotypes.
Viruses    July 26, 2023   Volume 15, Issue 8 1626 doi: 10.3390/v15081626
Carossino M, Balasuriya UBR, Thieulent CJ, Barrandeguy ME, Vissani MA, Parreño V.Equine rotavirus A (ERVA) is the leading cause of diarrhea in foals, with G3P[12] and G14P[12] genotypes being the most prevalent. Recently, equine G3-like RVA was recognized as an emerging infection in children, and a group B equine rotavirus (ERVB) was identified as an emergent cause of foal diarrhea in the US. Thus, there is a need to adapt molecular diagnostic tools for improved detection and surveillance to identify emerging strains, understand their molecular epidemiology, and inform future vaccine development. We developed a quadruplex TaqMan RT-qPCR assay for differentiation of ERVA an...
Unilateral keratitis secondary to Leishmania spp. infection in a horse: Clinical signs and successful topical therapy.
Veterinary ophthalmology    July 25, 2023   doi: 10.1111/vop.13134
Matas Riera M, Cardenas Nadal M, Martínez-Sogues L, Ferrer L.Dermatological clinical signs have been seldom reported in the literature secondary to equine leishmaniasis. This case depicts the clinical signs, treatment, and outcome of a young horse with a pink, elevated lesion on the ventromedial quadrant of the cornea. A corneal cytology was performed and revealed the presence of leishmania amastigotes reaching the diagnosis of keratitis secondary to leishmania. Surgical resection was recommended but the owner declined the procedure, and the lesion was treated with a topical antimonial for 6 weeks. The lesion reduced remarkably during the first weeks ...
Identification of vaccine candidates against rhodococcus equi by combining pangenome analysis with a reverse vaccinology approach.
Heliyon    July 25, 2023   Volume 9, Issue 8 e18623 doi: 10.1016/j.heliyon.2023.e18623
Liu L, Yu W, Cai K, Ma S, Wang Y, Ma Y, Zhao H. () is a zoonotic opportunistic pathogen that can cause life-threatening infections. The rapid evolution of multidrug-resistant and the fact that there is no currently licensed effective vaccine against warrant the need for vaccine development. Reverse vaccinology (RV), which involves screening a pathogen's entire genome and proteome using various web-based prediction tools, is considered one of the most effective approaches for identifying vaccine candidates. Here, we performed a pangenome analysis to determine the core proteins of . We then used the RV approach to examine the subcellular l...
First Insight into the Seroepidemiology of Hepatitis E Virus (HEV) in Dogs, Cats, Horses, Cattle, Sheep, and Goats from Bulgaria.
Viruses    July 21, 2023   Volume 15, Issue 7 doi: 10.3390/v15071594
Tsachev I, Gospodinova K, Pepovich R, Takova K, Kundurzhiev T, Zahmanova G, Kaneva K, Baymakova M.In recent years, hepatitis E virus (HEV) infection has been found to be widespread among different animal species worldwide. In Bulgaria, high HEV seropositivity was found among pigs (60.3%), wild boars (40.8%), and East Balkan swine (82.5%). The aim of the present study was to establish the seroprevalence of HEV among dogs, cats, horses, cattle, sheep, and goats in Bulgaria. In total, 720 serum samples from six animal species were randomly collected: dogs-90 samples; cats-90; horses-180; cattle-180; sheep-90; and goats-90. The serum samples were collected from seven districts of the country: ...
Detection of Viable Streptococcus equi equi Using Propidium Monoazide Polymerase Chain Reaction.
Journal of equine veterinary science    July 20, 2023   Volume 128 104893 doi: 10.1016/j.jevs.2023.104893
Boyle AG, O'Shea K, Stefanovski D, Rankin SC.There is debate around the clinical significance of Streptococcus equi subsp. equi detection in low numbers using quantitative real-time PCR (qPCR). Propidium monoazide (PMA) qPCR has been used to differentiate DNA from viable and nonviable bacterial cells. The aim of this study was to evaluate the ability of PMA eqbE SEQ2190 triplex qPCR to differentiate DNA from viable and nonviable S. equi in positive and suspect positive clinical specimens. Fifty-seven stored (frozen and refrigerated) positive (36) or suspect positive (21) clinical specimens (determined via SeeI qPCR as the gold standard) ...
Equine bronchial epithelial cells are susceptible to cell entry with a SARS-CoV-2 pseudovirus but reveal low replication efficiency.
American journal of veterinary research    July 18, 2023   1-11 doi: 10.2460/ajvr.23.06.0132
Legere RM, Allegro AR, Affram Y, Silveira BPD, Fridley JL, Wells KM, Oezguen N, Burghardt RC, Wright GA, Pollet J, Bordin AI, Figueiredo P....To examine the susceptibility of cultured primary equine bronchial epithelial cells (EBECs) to a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus relative to human bronchial epithelial cells (HBECs). Methods: Primary EBEC cultures established from healthy adult horses and commercially sourced human bronchial epithelial cells (HBECs) were used as a positive control. Methods: Angiotensin-converting enzyme 2 (ACE2) expression by EBECs was demonstrated using immunofluorescence, western immunoblot, and flow cytometry. EBECs were transduced with a lentivirus pseudotyped with ...
Henipavirus zoonosis: outbreaks, animal hosts and potential new emergence.
Frontiers in microbiology    July 17, 2023   Volume 14 1167085 doi: 10.3389/fmicb.2023.1167085
Li H, Kim JV, Pickering BS.Hendra virus (HeV) and Nipah virus (NiV) are biosafety level 4 zoonotic pathogens causing severe and often fatal neurological and respiratory disease. These agents have been recognized by the World Health Organization as top priority pathogens expected to result in severe future outbreaks. HeV has caused sporadic infections in horses and a small number of human cases in Australia since 1994. The NiV Malaysia genotype (NiV-M) was responsible for the 1998-1999 epizootic outbreak in pigs with spillover to humans in Malaysia and Singapore. Since 2001, the NiV Bangladesh genotype (NiV-B) has been t...
JMM Profile: West Nile virus.
Journal of medical microbiology    July 17, 2023   Volume 72, Issue 7 doi: 10.1099/jmm.0.001730
Sewgobind S, McCracken F, Schilling M.West Nile virus (WNV) is a positive-sense single-stranded RNA virus belonging to the Flaviviridae family and is maintained in an enzootic cycle between avian hosts and mosquito vectors. Humans, horses and other mammals are susceptible to infection but are dead-end hosts due to a low viraemia. The disease can manifest itself in a variety of clinical signs and symptoms in people and horses from mild fever to severe encephalitis and morbidity. There are no vaccines licensed for human protection, but parts of Europe, North America, Africa and Australia have vaccines commercially available for hors...
Equine synovial sepsis laboratory submissions yield a low rate of positive bacterial culture and a high prevalence of antimicrobial resistance.
American journal of veterinary research    July 17, 2023   Volume 84, Issue 8 doi: 10.2460/ajvr.23.05.0085
Pearson GB, Papa B, Mosaddegh A, Cooper H, Aprea M, Pigott J, Altier C, Cazer CL, Reesink HL.To investigate (1) variables associated with the likelihood of obtaining a positive culture, (2) commonly isolated microorganisms, and (3) antimicrobial resistance patterns of isolates from horses with presumptive synovial sepsis. Methods: Synovial fluid, synovium, and bone samples from equine cases with presumptive synovial sepsis submitted to the Cornell University Animal Health Diagnostic Center from 2000 to 2020 for microbial culture and antimicrobial sensitivity testing. Methods: Univariable and multivariable analyses were performed to determine the effect of variables on the likelihood o...
Diversity of the Bacterial and Viral Communities in the Tropical Horse Tick, Dermacentor nitens, in Colombia.
Pathogens (Basel, Switzerland)    July 16, 2023   Volume 12, Issue 7 doi: 10.3390/pathogens12070942
Holguin-Rocha AF, Calle-Tobon A, Vásquez GM, Astete H, Fisher ML, Tobon-Castano A, Velez-Tobon G, Maldonado-Ruiz LP, Silver K, Park Y....Ticks are obligatory hematophagous ectoparasites that transmit pathogens among various vertebrates, including humans. The microbial and viral communities of ticks, including pathogenic microorganisms, are known to be highly diverse. However, the factors driving this diversity are not well understood. The tropical horse tick, Dermacentor nitens, is distributed throughout the Americas and it is recognized as a natural vector of Babesia caballi and Theileria equi, the causal agents of equine piroplasmosis. In this study, we characterized the bacterial and viral communities associated with partial...
Monitoring the epidemic of West Nile virus in equids in Spain, 2020-2021.
Preventive veterinary medicine    July 13, 2023   Volume 217 105975 doi: 10.1016/j.prevetmed.2023.105975
Gonzálvez M, Franco JJ, Barbero-Moyano J, Caballero-Gómez J, Ruano MJ, Martínez R, Cano-Terriza D, García-Bocanegra I.The largest epidemic of West Nile virus (WNV) reported ever in Spain in both humans and equines occurred in 2020, affecting 77 humans and 139 equine herds. Here, we aimed to monitor the outbreaks detected in equid herds in Andalusia (southern Spain), the Spanish region where 89.9% of the outbreaks were reported, and to evaluate the virus circulation and risk factor associated with WNV exposure in the affected herds. The first WNV case was detected in mid-July 2020, the number of outbreaks peaked in mid-August and the last one was confirmed on 26th October 2020. WNV lineage 1 was detected in 12...
Is leishmaniasis the new emerging zoonosis in the world?
Veterinary research communications    July 12, 2023   doi: 10.1007/s11259-023-10171-5
Montaner-Angoiti E, Llobat L.Leishmania is a genus of parasitic protozoa that causes a disease called leishmaniasis. Leishmaniasis is transmitted to humans through the bites of infected female sandflies. There are several different species of Leishmania that can cause various forms of the disease, and the symptoms can range from mild to severe, depending on species of Leishmania involved and the immune response of the host. Leishmania parasites have a variety of reservoirs, including humans, domestic animals, horses, rodents, wild animals, birds, and reptiles. Leishmaniasis is endemic of 90 countries, mainly in South Amer...
Borna disease virus 1 infection in alpacas: Comparison of pathological lesions and viral distribution to other dead-end hosts.
Veterinary pathology    July 11, 2023   3009858231185107 doi: 10.1177/03009858231185107
Fürstenau J, Richter MT, Erickson NA, Große R, Müller KE, Nobach D, Herden C, Rubbenstroth D, Mundhenk L.Borna disease is a progressive meningoencephalitis caused by spillover of the Borna disease virus 1 (BoDV-1) to horses and sheep and has gained attention due to its zoonotic potential. New World camelids are also highly susceptible to the disease; however, a comprehensive description of the pathological lesions and viral distribution is lacking for these hosts. Here, the authors describe the distribution and severity of inflammatory lesions in alpacas ( = 6) naturally affected by this disease in comparison to horses ( = 8) as known spillover hosts. In addition, the tissue and cellular distribu...
High Seroprevalence of Lawsonia intracellularis in Thoroughbred Farms in Southern Brazil.
Journal of equine veterinary science    July 7, 2023   Volume 128 104890 doi: 10.1016/j.jevs.2023.104890
de Souza RP, Mousquer MA, Müller V, Barbosa JCR, Leite FPL, Guedes RMC, Curcio BDR, Nogueira CEW.The aim of the present study was to carry out a serological survey to identify the seroprevalence of Lawsonia intracellularis in six Thoroughbred farms in the Southern region of the state of Rio Grande do Sul, Brazil. During 2019 and 2020, blood samples from 686 Thoroughbred horses were obtained from six different breeding farms. Horses were divided into groups according to age: (1) broodmares (>5 years), (2) two-year-old foals, (3) yearlings, and (4) 0-6 months-old foals. Blood samples were collected by venipuncture of the external jugular vein. The detection of antibodies (IgG) against L. in...
Nontuberculous Mycobacteria in Horses: A Narrative Review.
Veterinary sciences    July 6, 2023   Volume 10, Issue 7 doi: 10.3390/vetsci10070442
Li L, Maboni G, Lack A, Gomez DE.Nontuberculous mycobacteria (NTM) infections are increasing in human and veterinary medicine. Although horses were initially thought to be resistant to NTM infection, reports of horses suffering from gastrointestinal, respiratory, and reproductive diseases associated with NTM have increased in the last few decades. The aim of this literature review is to summarize the mycobacteria species found in horses, describe clinical manifestations, diagnostic and treatment approaches, and public health concerns of NTM infection in horses. Clinical manifestations of NTM in horses include pulmonary diseas...
Host cell restriction factors of equine infectious anemia virus.
Virologica Sinica    July 5, 2023   Volume 38, Issue 4 485-496 doi: 10.1016/j.virs.2023.07.001
Wang XF, Zhang X, Ma W, Li J, Wang X.Equine infectious anemia virus (EIAV) is a member of the lentivirus genus in the Retroviridae family and is considered an animal model for HIV/AIDS research. An attenuated EIAV vaccine, which was successfully developed in the 1970s by classical serial passage techniques, is the first and only lentivirus vaccine that has been widely used to date. Restriction factors are cellular proteins that provide an early line of defense against viral replication and spread by interfering with various critical steps in the viral replication cycle. However, viruses have evolved specific mechanisms to overcom...
Epidemiological study on equine coccidiosis in North and Northeast of Iran.
Veterinary medicine and science    July 4, 2023   doi: 10.1002/vms3.1197
Gorji FF, Sadr S, Borji H.Eimeria is a genus of protozoan parasites that infect many animal species, including horses. We conducted a cross-sectional study of indigenous breeds of horses from the North and Northeast of Iran to establish the prevalence and distribution of Eimeria species. Methods: Using standard coprological techniques, 340 faecal samples from randomly selected horses (141 from North Iran and 199 from Northeast Iran) were examined for Eimeria oocyst. Results: Out of 340 samples, only three from north Iran were positive for coccidiosis. Infections occurred by Eimeria leuckarti. The mean intensity of oocy...
Genomic Analysis of Clostridioides difficile Recovered from Horses in Western Australia.
Microorganisms    July 3, 2023   Volume 11, Issue 7 doi: 10.3390/microorganisms11071743
Hain-Saunders NMR, Knight DR, Bruce M, Byrne D, Riley TV.Clostridioides difficile poses an ongoing threat as a cause of gastrointestinal disease in humans and animals. Traditionally considered a human healthcare-related disease, increases in community-associated C. difficile infection (CDI) and growing evidence of inter-species transmission suggest a wider perspective is required for CDI control. In horses, C. difficile is a major cause of diarrhoea and life-threatening colitis. This study aimed to better understand the epidemiology of CDI in Australian horses and provide insights into the relationships between horse, human and environmental strains...
Prevalence of equine parvovirus-hepatitis in healthy broodmares in Ontario, Canada.
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire    July 3, 2023   Volume 87, Issue 3 169-175 doi: 10.25318/3210037301-eng
Papapetrou MA, Arroyo LG, Meister TL, Baird JD, Steinmann E, Lillie BN.Equine parvovirus-hepatitis (EqPV-H) was first reported from the serum and liver tissue of a horse diagnosed with Theiler's disease in the United States in 2018. Theiler's disease, also known as equine serum hepatitis, is a severe hepatitis with fulminant hepatic necrosis. The disease has most frequently been reported following the administration of equine-origin biological products; however, it has also been reported in in-contact horses with no prior biologic administration. EqPV-H has been detected in clinically healthy horses in North America (USA, Canada), Europe (Germany, Austria, Sloven...
Venezuelan Equine Encephalitis
   July 3, 2023  
Crosby B, Crespo ME.Venezuelan Equine Encephalitis Virus (VEEV) is the causative viral pathogen of Venezuelan Equine Encephalitis (VEE). Outbreaks frequently involve both equines – including horses, donkeys, mules, zebras – and humans. Outbreaks may range over a large geographic area and may last for several months to years. Sporadic epidemic outbreaks occur most commonly in Central and South America. VEEV exists as both a natural pathogen and a laboratory-developed biologic weapon. Outbreaks have been reported in several South and Central American countries, including Venezuela, Colombia, Peru, Ecuador, C...
Detection of Equus caballus Papillomavirus in Equine Aural Plaque Samples.
Journal of equine veterinary science    July 1, 2023   Volume 128 104877 doi: 10.1016/j.jevs.2023.104877
Bromberger CR, Costa JR, Herman M, Hernandez JM, Albertino LG, Alves CEF, Borges AS, Oliveira-Filho JP.Aural plaques have been linked to Equus caballus papillomavirus (EcPV). Ten types of EcPVs have already been described; however, only EcPVs 1, 3, 4, 5, and 6 have been observed in association with aural plaques. Accordingly, the objective of this study was to evaluate the presence of EcPVs in equine aural plaque samples. A total of 29 aural plaque samples (from 15 horses) were collected and assessed for the presence of the DNA of these EcPVs by PCR. Additionally, 108 aural plaque samples used in previous research were evaluated for the presence of EcPVs 8 and 9. Previously described primers we...
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