Zum Studium des prophylaktischen Einflusses ausgewählter antibiotischer Leistungsförderer [Avilamycin (10 ppm), Avoparcin (15 ppm), Virginiamycin (20 ppm)] und ionophorer Antikokzidia [Monensin (100 ppm), Narasin (70 ppm)] sowie des metaphylaktischen bzw. therapeutischen Einsatzes von Tylosin [Tylan 0,5 g/l H2O] auf das Auftreten und die Ausprägung der Clostridium (Cl.) perfringens-Enterotoxämie (CPE) wurden Untersuchungen an insgesamt 33 Versuchsgruppen mit 825 Broilerküken durchgeführt. Die Erkrankung konnte mittels intraduodenaler Inokulation einer Vollkultur Cl. perfringens Typ A (ATCC 3624) sicher reproduziert werden. Die Morbiditätsrate betrug in allen infizierten Gruppen 100 %. An klinischer Symptomatik zeigte sich hauptsächlich profuser wässriger Durchfall. Schwerere Störungen, Apathie und Anorexie, waren selten und in allen beobachteten Fällen vom schnellen Tod des betreffenden Tieres begleitet. Allgemein fiel auf, daß die infizierten und nicht medikamentierten Tiere schneller und länger erkrankten. Bei infizierten und unmedikamentierten Tieren ergab sich eine Mortalitätsrate von 16 bis 36 %, in den medikamentierten Gruppen maximal 8 %. Tylosin zeigte eine sehr gute metaphylaktischen bzw. therapeutische Wirkung. Die Lebendmasseentwicklung betrachtend, konnte Avoparcin unter den Leistungsförderern beste Ergebnisse erzielen. Ähnliche Resultate wurden in den Kombinationsgruppen [Avilamycin plus Monensin oder Narasin] bzw. mittels Narasin erreicht. In absteigender Reihenfolge zeigten Avilamycin, Virginiamycin und Monensin eine geringere leistungsfördernde Wirkung. Die Bestimmung fäkaler bzw. ileozäkaler Clostridienkonzentrationen lebender, respektive verendeter Hühner erbrachte nur wenige und relativ unbedeutende statistisch gesicherte Korrelationen zu anderen Ergebnissen. Es konnten keine Zusammenhänge zwischen Erregerzahl und Lebendmassezunahme, bzw. Todesursache, aufgedeckt werden. Die Resultate aus allen Versuchen zusammenfassend, müssen den Kombinationen von Avilamycin mit Narasin bzw. Monensin beste Effekte hinsichtlich einer positiven Beeinflussung CPE-bedingter Morbidität, Mortalität und Lebendmasseverluste bescheinigt werden. Tylosin war in der Lage, die Verlustzahlen durch CPE rasch zu senken. Für die Ausprägung kompensatorischer Effekte hinsichtlich der Lebendmasseverluste unter der Infektion muß mit einer größeren Zeitspanne gerechnet werden. Die Polyether und auch Avilamycin sind als Futtermittelzusatzstoffe für die europäische Geflügelhaltung zugelassen. Durch die ständige Kokzidiosebedrohung in den Hühnerbeständen kann auf einen prophylaktischen Einsatz antikokzidieller Futtermittelzusatzstoffe momentan nicht verzichtet werden. Es ist zu vermuten, daß es durch den simultanen Einsatz von Polyether und Leistungsförderer zu einer positiven Beeinflussung der schädigenden Wechselwirkungen von Kokzidien und Cl. perfringens im Darm kommt. Bei vorhandener Empfindlichkeit der Eimerien sollte dies sowohl die Bekämpfung von CPE als auch von Kokzidiosen begünstigen. Der positive Eindruck von Avoparcin spielt, bedingt durch das europaweite Verbot, momentan für die Praxis keine Rolle. Die Entwicklungstendenzen auf dem Sektor antibiotisch wirksamer Futtermittelzusatzstoffe, eng verknüpft mit der bakteriellen Resistenzproblematik, werden in der Arbeit ausführlich diskutiert. / Investigations with 825 chickens in 33 trials were performed in order to find out the prophylactic effect of selected antibiotic growth promoters [avilamycin (10 ppm), avoparcin (15 ppm) virginiamycin (20 ppm)] and polyether ionophore antibiotics [monensin (100 ppm), narasin (70 ppm)] on the incidence of Clostridium (Cl.) perfringens enterotoxemia (CPE) in chickens as well as the therapeutic resp. metaphylactic influence of tylosin [Tylan 0,5 g/l H20]. The enterotoxemia could be reproduced regularly by intraduodenal infection with high numbers of vegetative cells of Cl. perfringens type A (ATCC 3624). The morbidity rate always reached 100 %. In spite of a profuse and watery diarrhoea the chickens normally showed no further considerable disturbances of the general status. Apathy or anorexia were rather rare and immediately followed by Exitus letalis of the related chickens. It was striking that the infected and non-medicated broilers contracted the disease more quickly and for a longer time. The mortality rate among the infected and non-medicated animals was 16 to 36 %, among the medicated groups max. 8 %. Tylosin showed a considerable metaphylactic effect in decreasing CPE mortality. The avoparcin group showed the best weight gain among the growth promoters, comparable to the results by means of the combinations [avilamycin + monensin or narasin] or narasin only. Decreasingly avilamycin, virginiamycin and monensin were less successful. Analysing the faecal resp. ileocecal quantities of Cl. perfringens adduced only a few statistically guaranteed correlation with other results. There was no causal connection between numbers of Cl. perfringens and life weight development. It was impossible to discover a numerical threshold of germs responsible for the death of the chickens. Summarising all the results of the entire attempts the combinations of avilamycin and narasin resp. monensin were the most effective concerning the reduction of morbidity, mortality and life weight losses by CPE. By application of tylosin it was possible to stop the mortality rate quickly. But it needs more time to achieve reductions of the CPE related weight losses. The two polyethers and also avilamycin are still admitted in the European Union. Currently an abandonment of anticoccidial feed supplements seems to be impossible due to the present danger of coccidiosis in poultry. By means of monensin/narasin plus avilamycin the adverse health effects of interactions of both pathogens should be reduced. Presupposing susceptibility of the coccida this should be a notable contribution to a better controlling and to the prevention of CPE and coccidiosis, too.

Einleitung: Im Rahmen umfangreicher statistischer Erhebungen in den Jahren 2007 bis 2017 wurde der Gesundheitsstatus von konventionell und ökologisch gemästeten Puten während der Aufzucht- bzw. Mastphase in Deutschland erfasst. In der unmittelbar vorangegangenen Studie (Bio1) wurde festgestellt, dass ökologisch gemästete Kelly Broad Breasted Bronze (BBB)-Puten im Vergleich zu konventionell gemästeten B.U.T. 6-Puten eine signifikant erhöhte Prävalenz für Leberveränderungen aufwiesen. Besonders auffällig war ein um das Neunfache erhöhtes Auftreten grün gefärbter Lebern (GL). Ziele der Untersuchungen: Ziel der vorliegenden Forschungsarbeit war die explorative Erhebung und Evaluation infektiöser Faktoren, welche einen kausalen Einfluss auf die Entstehung einer Grünfärbung der Leber bei nach ökologischen Richtlinien gehaltenen Mastputen haben. Tiere, Material und Methoden: Insgesamt wurden 343 Bronze-Hennen einer Cartier Genetik aus fünf verschiedenen Mastbeständen in jeweils zwei Durchgängen mit je zwei Untersuchungsterminen (U1 = 70.– 75. Masttag, U2 = 120. – 127. Masttag) klinisch und pathologisch-anatomisch untersucht. Die Stichprobengröße von 20 Puten je Untersuchungstermin errechnete sich anhand einer erwarteten Prävalenz für GL von 27,7 % mit einem unteren 95 %-Konfidenzintervall von 18,5 % (ausgehend von Bio1), sowie einer laut VO (EG) Nr. 889/2008 maximal erlaubten Herdengröße von 2500 Tieren. Bei jeweils sechs Hennen ohne und, falls GL vorhanden, bis zu sechs Hennen mit GL schließen sich pathologisch-histologische, mikrobiologische, parasitologische und virologische Untersuchungen an. Insgesamt wurden 130 Hennen weiterführend untersucht. Die statistischen Berechnungen wurden je nach Skalierung, Verteilung und Homoskedastizität der Daten als Korrelationsberechnungen (Korrelationskoeffizient nach Bravais-Pearson, Spearman'scher Rangkorrelationskoeffizient) und Hypothesentests (Chi-Quadrat-Test, ungepaarter t-Test, Mann-Whitney-U-Test) durchgeführt. Im Vergleich mehrerer Gruppen wurde eine einfaktorielle ANOVA bzw. der Kruskal-Wallis-Test mit sich jeweils anschließendem Tukey Post-Hoc Test oder Games-Howell Post-Hoc Test implementiert. Das Signifikanzniveau lag bei p ≤ 0,05. Ergebnisse: Die Prävalenzen für GL haben sich im Vergleich zur Vorstudie deutlich verringert. In Bio1 konnte dieser Befund bei 33,1 % der ökologisch gehaltenen Kelly-BBB-Hennen am Schlachthof erhoben werden. In der aktuellen Studie lag die Prävalenz bei den Bronze-Hennen einer Cartier-Genetik im Zuge der pathologisch-anatomischen Untersuchungen mit 9,0 % (95 %-KI [0,06;0,12]) deutlich darunter. Zwischen den einzelnen Beständen fanden sich jedoch Schwankungen zwischen 0,0 % bis 68,4 %. Pathomorphologisch zeigten die GL im Vergleich zu physiologischen Lebern signifikant häufiger eine Infiltration von Entzündungszellen (U1 p ≤ 0,05, U2 p ≤ 0,05) und waren signifikant schwerer (U1 p ≤ 0,05, U2 p ≤ 0,01). Hennen mit GL wiesen zu beiden Untersuchungszeitpunkten ein signifikant geringeres Körpergewicht auf (U1 p ≤ 0,05, U2 p ≤ 0,001). Zu U1 waren bei Hennen mit einer GL signifikant häufiger entzündliche Reaktionen im Dünndarm festzustellen (p ≤ 0,001). Zu U2 wiesen Hennen mit GL signifikant häufiger makroskopische (p ≤ 0,001) und histologische (p ≤ 0,05) Knochenläsionen auf. Die kausale Pathogenese unterscheidet sich zwischen den zwei Untersuchungszeitpunkten. Der Nachweis des immunsupprimierenden Hämorrhagischen Enteritis-Virus (HEV) korreliert signifikant mit der GL in der frühen Mastphase (p ≤ 0,001). Weiterhin wiesen die Hennen mit HEV-Infektion signifikant häufiger pathologische Veränderungen in Milz (Splenomegalie zu U2 p ≤ 0,001) und Duodenum (katarrhalische Duodenitis zu U1 p ≤ 0,001) auf. Zu U2 manifestierte sich bei Hennen mit GL eine signifikant reduzierte Gewichtsentwicklung (p ≤ 0,001). Die HEV positiven ungeimpften Hennen zeigten die höchsten Prävalenzen für GL zu beiden Untersuchungen (U1 = 35,9 %, U2 = 20,0 %), sowie die höchste mittlere Mortalitätsrate (9,15 %). Zu U1 konnten bei diesen Hennen die höchsten relativen Milzgewichte (0,12 %) und die für das HEV charakteristischen Einschlusskörper in Milz und Duodenum gefunden werden. Darüber hinaus wiesen diese Hennen zu U2 die höchsten Prävalenzen für pathologische Veränderungen am Bewegungsapparat (20,0 %) auf. Zu U2 korrelierten die GL signifikant mit dem Vorhandensein von makroskopischen Läsionen des Bewegungsapparats (p ≤ 0,001) sowie einer aseptischen Osteomyelitis (p ≤ 0,05). Somit kann bei den älteren Puten in wesentlichen Punkten vom klassischen Turkey Osteomyelitis Complex (TOC) und einem akuten Entzündungsgeschehen gesprochen werden. Der TOC charakterisiert sich dabei durch das gleichzeitige Vorliegen einer GL und pathologischen Prozessen am Bewegungsapparat, welche durch Escherichia coli und Staphylococcus aureus hervorgerufen werden können. Letzterer ließ sich bei einer Henne mit GL aus multiplen Organen und dem Kniegelenk isolieren. Schlussfolgerungen: Die Befunde dieser Forschungsarbeit sprechen für eine deutliche gesundheitliche Beeinträchtigung der Puten mit einer GL. Die Schädigung der Leber als zentrales Stoffwechselorgan stellt dabei einen ernstzunehmenden Indikator für die Tiergesundheit bei Mastputen dar. Des Weiteren lassen die Befunde vermuten, dass eine Infektion mit dem HEV die Tiergesundheit auch langfristig negativ beeinflusst. Liegen innerhalb eines Durchgangs Prävalenzen von mehr als 1 % für GL vor, so sollte im Sinne des Tierwohls und des Verbraucherschutzes eine Ursachenanalyse mit folgenden Reduktionsmaßnahmen durchgeführt werden. Die Schwankungen der Prävalenzen zwischen den einzelnen Beständen sprechen jedoch dafür, dass die Entstehung der GL keine universell in der ökologischen Putenhaltung vorkommende Problematik darstellt. Der mit den GL im Zusammenhang stehende TOC konnte in dieser Studie als ein multifaktoriell bedingtes Geschehen charakterisiert werden. Dieses wird unter anderem von interagierenden Faktoren wie der Gesundheitsprophylaxe (insb. die Impfung gegen HEV), bakteriellen, parasitären oder viralen Erregern beeinflusst. In Bezug auf das HEV konnte ein negativer Effekt auf die Tiergesundheit inklusive häufig grün gefärbter Lebern nachgewiesen werden. Somit stellt die HEV-Impfung mit einer an die aktuell zirkulierenden Stämme angepassten Vakzine eine Möglichkeit dar diesen Faktoren auch langfristig vorzubeugen.

Rotavirus infection causes severe gastroenteritis, affecting all children under the age of five regardless of hygiene or water quality. The currently licensed vaccines succeeded in reducing diarrhoea worldwide, but they still have shortcomings, especially the efficacy of the vaccines in developing countries. One of the main reasons for this can be due to the difference in strains, since the strains used to develop the currently licensed vaccines (RotaTeq and Rotarix) were selected from strains circulating in the developed world (G1, G2, G3 and G4), while the main strains present in Africa (G8, G9 and G12) were not included. A second shortcoming of the currently licensed vaccines is the cost of these vaccines. The vaccines are very expensive and most developing countries cannot afford the vaccines as well as the fact that the manufacturing companies cannot produce enough vaccines for all the countries. An attractive alternative to the currently licensed rotavirus vaccines is the non-live vaccine candidate, virus-like particles, which can provide a possible cheaper, safer and efficacious alternative or complement the currently licensed vaccines. Therefore, in this study a South African G9P[6] rotavirus strain, RVA/Humanwt/ ZAF/GR10924/1999/G9P[6], was used to determine whether or not co-expression of the structural proteins VP2 (genome segment 2) and VP6 (genome segment 6) was possible in bacteria and yeast. The South African GR10924 G9P[6] neonatal strain was previously obtained from a stool sample and the nucleotide consensus sequence was determined for both genome segment 2 (VP2) and genome segment 6 (VP6). Bacterial codon optimised coding regions or open reading frames were used in this study. The open reading frames (ORFs) of the genome segments encoding, VP2 and VP6, were cloned into the expression vector pETDuet-1, which allows for the simultaneous expression of two genes in bacteria. The ORF of genome segment 6 was purchased from GeneScript and the ORF of genome segment 2 was obtained from Dr AC Potgieter (Deltamune (Pty) Ltd R&D, South Africa). Compatible restriction enzyme sites were used to sub-clone the ORF of the bacterial codon optimised genome segments into the expression vector. Only the expression of the VP6 protein in bacteria was observed with Coomassie stained SDS-PAGE. The ORFs encoding VP2 (genome segment 2) and VP6 (genome segment 6) of the wild type GR10924 G9P[6] strain were cloned into the wide range yeast expression system vector, pKM173, which allows for the simultaneous expression of more than one gene. Several yeast strains were used in this study namely Kluyveromyces marxianus, Kluyveromyces lactis, Candida deformans, Saccharomyces cerevisiae, Yarrowia lipolytica, Arxula adeninivorans, Hansenula polymorpha and Debaryomyces hansenii. Expression of both proteins was not detected in the several yeast strains, as seen with western blot analysis. DNA extractions were done on two colonies of each yeast strain that were used for western blot analysis to evaluate successful integration into the yeast genomes. Only a few of the colonies contained either both of the genome segments or only one of the two genome segments of interest. To summarise, the simultaneous expression of VP2 and VP6 from rotavirus GR10924 G9P[6] was not successful in bacteria or yeast, but it was possible to soluble express the bacterial codon optimised GR10924 G9P[6] VP6 in bacteria using the pETDuet-1 as expression vector. / MSc (Biochemistry), North-West University, Potchefstroom Campus, 2015

Rotavirus infection causes severe gastroenteritis, affecting all children under the age of five regardless of hygiene or water quality. The currently licensed vaccines succeeded in reducing diarrhoea worldwide, but they still have shortcomings, especially the efficacy of the vaccines in developing countries. One of the main reasons for this can be due to the difference in strains, since the strains used to develop the currently licensed vaccines (RotaTeq and Rotarix) were selected from strains circulating in the developed world (G1, G2, G3 and G4), while the main strains present in Africa (G8, G9 and G12) were not included. A second shortcoming of the currently licensed vaccines is the cost of these vaccines. The vaccines are very expensive and most developing countries cannot afford the vaccines as well as the fact that the manufacturing companies cannot produce enough vaccines for all the countries. An attractive alternative to the currently licensed rotavirus vaccines is the non-live vaccine candidate, virus-like particles, which can provide a possible cheaper, safer and efficacious alternative or complement the currently licensed vaccines. Therefore, in this study a South African G9P[6] rotavirus strain, RVA/Humanwt/ ZAF/GR10924/1999/G9P[6], was used to determine whether or not co-expression of the structural proteins VP2 (genome segment 2) and VP6 (genome segment 6) was possible in bacteria and yeast. The South African GR10924 G9P[6] neonatal strain was previously obtained from a stool sample and the nucleotide consensus sequence was determined for both genome segment 2 (VP2) and genome segment 6 (VP6). Bacterial codon optimised coding regions or open reading frames were used in this study. The open reading frames (ORFs) of the genome segments encoding, VP2 and VP6, were cloned into the expression vector pETDuet-1, which allows for the simultaneous expression of two genes in bacteria. The ORF of genome segment 6 was purchased from GeneScript and the ORF of genome segment 2 was obtained from Dr AC Potgieter (Deltamune (Pty) Ltd R&D, South Africa). Compatible restriction enzyme sites were used to sub-clone the ORF of the bacterial codon optimised genome segments into the expression vector. Only the expression of the VP6 protein in bacteria was observed with Coomassie stained SDS-PAGE. The ORFs encoding VP2 (genome segment 2) and VP6 (genome segment 6) of the wild type GR10924 G9P[6] strain were cloned into the wide range yeast expression system vector, pKM173, which allows for the simultaneous expression of more than one gene. Several yeast strains were used in this study namely Kluyveromyces marxianus, Kluyveromyces lactis, Candida deformans, Saccharomyces cerevisiae, Yarrowia lipolytica, Arxula adeninivorans, Hansenula polymorpha and Debaryomyces hansenii. Expression of both proteins was not detected in the several yeast strains, as seen with western blot analysis. DNA extractions were done on two colonies of each yeast strain that were used for western blot analysis to evaluate successful integration into the yeast genomes. Only a few of the colonies contained either both of the genome segments or only one of the two genome segments of interest. To summarise, the simultaneous expression of VP2 and VP6 from rotavirus GR10924 G9P[6] was not successful in bacteria or yeast, but it was possible to soluble express the bacterial codon optimised GR10924 G9P[6] VP6 in bacteria using the pETDuet-1 as expression vector. / MSc (Biochemistry), North-West University, Potchefstroom Campus, 2015

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

O coronavírus de perus (TCoV) é o agente etiológico associado a síndrome de mortalidade entérica das aves (PEMS). PEMS é uma enfermidade entérica, aguda e altamente contagiosa dos perus caracterizada por depressão, anorexia, diarréia e alta mortalidade em lotes de perus comerciais. A presença do coronavírus de perus (TCov) foi pesquisada em 29 amostras de conteúdo intestinal de perus entre 10 e 104 dias de idade que apresentaram enterite severa no período de 2004 a 2006. A detecção do TcoV foi realizada realizada através da técnica da transcriptase reversa e da reação em cadeia pela polimerase (RT-PCR), mediante a amplificação da região 3 UTR, seguida pela amplificação dos genes 3 e 5. A caracterização molecular dos vírus foi realizada mediante a amplificação dos genes 3 e 5, que mostrou similaridade genética entre as amostras, mas diferenças com as sequencias dos outros TCoVs publicados previamente. Em relação ao gene 3, as amostras apresentaram maior relação com o vírus da bronquite infecciosa das aves (IBV), enquanto que com o gene 5 houve maior identidade com os cronavírus de faisão (PhCoV). Nossos resultados sugerem que a estratégia de amplificação da região 3 UTR provou ser uma estratégia eficaz para a detecção do TcoV em conteúdo intestinal. / Turkey coronavirus (TCoV) is causative agent associated to Poult Enteritis and Mortality Syndrome (PEMS) in turkeys wideworld. The disease is characterized by an acute highly contagious enteric disease of turkeys characterized by depression, anorexia, diarrhea and high mortality in co mMercial turkey flocks. The presence of turkey coronavirus (TCoV) in 29 intestinal content samples from turkey flocks aged between 10 and 104 days with severe enteritis was monitored in the period of 2004 to 2006. TCoV detection was accomplished by the reverse transcriptase-polymerase chain reaction (RT-PCR), through amplification of the 3´UTR region, followed by amplification of genes 3 and 5. Molecular characterization of the viruses was done through amplification of genes 3 and 5, and showed evidence of genetic similarity between them, although they differed of sequences of other TCoVs described in the literature. In relation to gene 3, samples showed greater relationship with chicken infectious bronchitis virus (IBV), and while gene 5 showed greater identity with pheasant coronavirus (PhCoV). Our results suggest that the strategy of amplification of the 3´UTR region has proved to an effective means of detection of TCoV in intestinal contents.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

No description available.

O coronavírus de perus (TCoV) é o agente etiológico associado a síndrome de mortalidade entérica das aves (PEMS). PEMS é uma enfermidade entérica, aguda e altamente contagiosa dos perus caracterizada por depressão, anorexia, diarréia e alta mortalidade em lotes de perus comerciais. A presença do coronavírus de perus (TCov) foi pesquisada em 29 amostras de conteúdo intestinal de perus entre 10 e 104 dias de idade que apresentaram enterite severa no período de 2004 a 2006. A detecção do TcoV foi realizada realizada através da técnica da transcriptase reversa e da reação em cadeia pela polimerase (RT-PCR), mediante a amplificação da região 3 UTR, seguida pela amplificação dos genes 3 e 5. A caracterização molecular dos vírus foi realizada mediante a amplificação dos genes 3 e 5, que mostrou similaridade genética entre as amostras, mas diferenças com as sequencias dos outros TCoVs publicados previamente. Em relação ao gene 3, as amostras apresentaram maior relação com o vírus da bronquite infecciosa das aves (IBV), enquanto que com o gene 5 houve maior identidade com os cronavírus de faisão (PhCoV). Nossos resultados sugerem que a estratégia de amplificação da região 3 UTR provou ser uma estratégia eficaz para a detecção do TcoV em conteúdo intestinal. / Turkey coronavirus (TCoV) is causative agent associated to Poult Enteritis and Mortality Syndrome (PEMS) in turkeys wideworld. The disease is characterized by an acute highly contagious enteric disease of turkeys characterized by depression, anorexia, diarrhea and high mortality in co mMercial turkey flocks. The presence of turkey coronavirus (TCoV) in 29 intestinal content samples from turkey flocks aged between 10 and 104 days with severe enteritis was monitored in the period of 2004 to 2006. TCoV detection was accomplished by the reverse transcriptase-polymerase chain reaction (RT-PCR), through amplification of the 3´UTR region, followed by amplification of genes 3 and 5. Molecular characterization of the viruses was done through amplification of genes 3 and 5, and showed evidence of genetic similarity between them, although they differed of sequences of other TCoVs described in the literature. In relation to gene 3, samples showed greater relationship with chicken infectious bronchitis virus (IBV), and while gene 5 showed greater identity with pheasant coronavirus (PhCoV). Our results suggest that the strategy of amplification of the 3´UTR region has proved to an effective means of detection of TCoV in intestinal contents.

MSc (Microbiology) / Department of Microbiology / BACKGROUND: Human norovirus (NoV) is the etiological agent associated with acute gastroenteritis (AGE) in both children and adults worldwide. Children of <5 years of age, the elderly and individuals suffering from chronic diseases are potentially at high risk of NoV-associated illness. High morbidity and mortality rate associated with NoV have been reported worldwide. In children under the age of 5 years about 1.8 million death cases have been reported in developing countries alone. Despite the fact that the virus is affecting people of all age groups, there is lack of data to elucidate the importance and the role of NoV in children of the age above 5 years and adults. OBJECTIVE: To characterize human norovirus in patients with diarrhoea in rural communities of Vhembe district, Limpopo province. MATERIALS AND METHODS : From August 2017 to October 2018, outpatient between 5 and 68 years of age from rural communities of Vhembe district, Limpopo province were recruited for this study. A total of n=80 stool samples were collected from patients with diarrhoea and were kept at 4˚C throughout the transportation to the laboratory and refrigerated at - 20˚C prior to RNA extraction. Stool samples were tested for norovirus using the RIDA©GENE NOROVIRUS I & II real-time RT-PCR. The RNA extracts tested positive for norovirus were subjected to RT-PCR amplification. The RT-PCR products of the amplified fragments were sequenced, and phylogenetic trees were constructed by the neighbor-joining method using MEGA 7 software. RESULTS: NoV was detected in 13(16%) out of 80 stool samples collected, of which 6 (46%) strains belonged to norovirus GII and 7 (54%) strains to norovirus GI. A total of 5 genotypes were detected (GII.Pg, GII.1, GII.2, GII.4 Sydney 2012). The phylogenetic analysis revealed circulation of NoV genotypes with considerable diversity. CONCLUSION: This study illustrates NoV prevalence and substantial genetic diversity in patients above 5 years of age living in rural communities of Vhembe district, Limpopo province. Continued systematic surveillance to evaluate norovirus association with diarrhoea is needed to have a full picture on the epidemiology and disease burden in people of all the age groups. / NRF