Data_Sheet_1_Caecal Microbiota of Experimentally Campylobacter jejuni-Infected Chickens at Different Ages.docx

Campylobacter jejuni is the most common bacterial cause of foodborne zoonosis in the European Union. Infections are often linked to the consumption and handling of poultry meat. The aim of the present study was to investigate the caecal microbiota of birds infected with C. jejuni at different ages. Therefore, a total of 180 birds of the laying hybrid Lohmann Brown-Classic were housed in 12 subgroups of 15 animals each in three performed repetitions. Three birds per subgroup were experimentally infected with C. jejuni at an age of about 21 days and about 78 days (4.46 ± 0.35 log10 CFU/bird). Twenty-one days after experimental infection, microbiome studies were performed on 72 caecal samples of dissected birds (three primary infected and three further birds/subgroup). Amplification within the hypervariable region V 4 of the 16S rRNA gene was performed and sequenced with the Illumina MiSeq platform. Statistical analyses were performed using SAS® Enterprise Guide® (version 7.1) and R (version 3.5.2). Both factors, the experimental replication (p < 0.001) and the chickens’ age at infection (p < 0.001) contributed significantly to the differences in microbial composition of the caecal samples. The factor experimental replication explained 24% of the sample’s variability, whereas the factor age at infection explained 14% thereof. Twelve of 32 families showed a significantly different count profile between the two age groups, whereby strongest differences were seen for seven families, among them the family Campylobacteraceae (adjusted p = 0.003). The strongest difference between age groups was seen for a bacterial species that is assigned to the genus Turicibacter which in turn belongs to the family Erysipelotrichaceae (adjusted p < 0.0001). Correlation analyses revealed a common relationship in both chicken ages at infection between the absolute abundance of Campylobacteraceae and Alcaligenaceae, which consists of the genus Parasutterella. In general, concentrations of particular volatile fatty acids (VFA) demonstrated a negative correlation to absolute abundance of Campylobacteraceae, whereby the strongest link was seen for n-butyrate (−0.51141; p < 0.0001). Despite performing consecutive repetitions, the factor experimental replication contributed more to the differences of microbial composition in comparison to the factor age at infection.

欧共体内，空肠弯曲菌是引起食源性人畜共患病最常见的细菌病原体。感染通常与家禽肉的消费和处理有关。本研究旨在探讨不同年龄感染空肠弯曲菌的鸟类盲肠微生物群。因此，总共180只产蛋杂交种洛汉姆布朗-经典品种的鸟类被分为12个每组15只动物的子组，共进行了三次重复实验。在每个子组中，实验性地将3只约21天和约78天大的鸟类（每只鸟类4.46 ± 0.35 log10 CFU）感染空肠弯曲菌。实验感染后21天，对解剖的72只鸟类盲肠样本（每组中的三个原发性感染鸟类和三个其他鸟类）进行了微生物组研究。在16S rRNA基因高度可变区域V 4内进行了扩增，并使用Illumina MiSeq平台进行测序。统计分析使用SAS® Enterprise Guide®（版本7.1）和R（版本3.5.2）进行。实验重复（p < 0.001）和感染时的鸡龄（p < 0.001）这两个因素对盲肠样本微生物组成的差异有显著贡献。实验重复因素解释了样本变异性的24%，而感染时的年龄因素解释了14%。在32个家族中，有12个家族在两组年龄之间的计数轮廓存在显著差异，其中七个家族的差异最为显著，包括弯曲菌科（调整后的p = 0.003）。在年龄组之间最强的差异见于一种被归入属 Turicibacter 的细菌，该属属于欧氏菌科（调整后的p < 0.0001）。相关性分析揭示了在两种感染年龄的鸡中，Campylobacteraceae和Parasutterella属的Alcaligenaceae的绝对丰度之间存在共同的关系。总体而言，特定挥发性脂肪酸（VFA）的浓度与Campylobacteraceae的绝对丰度呈负相关，其中n-丁酸（−0.51141；p < 0.0001）的关联性最强。尽管进行了连续的重复实验，但实验重复因素对微生物组成差异的贡献大于感染时的年龄因素。